protease inhibition and detection

32
The cytoplasmic and nuclear proteasome complexes function to degrade ubiquitin-labeled proteins that are targeted for down regulation or disposal due to misfolding. Volume 4, Number 2 BIOFILES Life Science Protease Inhibition and Detection Broad Spectrum Protease Inhibitors Inhibitor Cocktails and Tablets Inhibitor Specificity Index for: Proteasome and Lysozomal Proteases Complement and Coagulation Factors Renin-Angiotensin System Enzymes Extracellular Matrix/MMPs Other Proteases Protease Detection: Kits and Substrates

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Page 1: Protease Inhibition and Detection

The cytoplasmic and nuclear proteasome complexes function to degrade ubiquitin-labeled proteins that are targeted for down regulation or disposal due to misfolding

Volume 4 Number 2

BioFilesLife Science

Protease Inhibition and Detection

Broad Spectrum Protease Inhibitors

Inhibitor Cocktails and Tablets

Inhibitor Specificity Index forProteasome and Lysozomal Proteases Complement and Coagulation FactorsRenin-Angiotensin System EnzymesExtracellular MatrixMMPsOther Proteases

Protease Detection Kits and Substrates

2

Life Science

BioFilesVolume 4 Number 2

Your gateway to Biochemicals and Reagents for Life Science Research from Sigmareg

BioFiles Online allows you to

bull Access any issue of BioFiles

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future issues

eBioFiles email notifications

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Register today for upcoming issues and eBioFiles announcements and receive a free IUBMB-Sigma-Nicholson Metabolic Pathway Chart sigmacombiofiles

Life Science TVFeaturing the Non-Specific Protease Activity Assay

The Sigmareg Life Science TV Web site is home to instructional informational and protocol based videos With regular updates to the library you will find videos covering many topics across the life science disciplines Currently available videos include

bull ATP Synthase Animation

bull GenomePlexreg Tutorial

bull TargeTronreg Technical Animation

The latest video addition to Life Science TV is titledNon-Specific Protease Activity Assay

This video displays a detailed protocol for determining the activity of proteases The universal assay procedure is useful for experiments ranging from sample preparation to QC analysis

View all Sigma Life Science Videos at sigma-aldrichcomlsvideos

Table of Contents

Introduction 3

Broad Spectrum Inhibitors of Proteolytic Enzyme Classes 4

Protease Inhibitor Panel 5

INHIB-1 Protease Inhibitor Panel Component Details 5

Protease Inhibitor Cocktails and Tablets 6

Protease Inhibitor Specificity Index 8

Product Listings 12

Aprotinin 15

E64 Analogs 20

Leupeptin 23

Trypsin Inhibitors 26

Protease Detection 29

Fluorescent Protease Detection 29

Non-Specific Protease Substrates 30

Technical content Christina Smith BS Robert Gates MS

Our Innovation Your Research mdash Shaping the Future of Life Science 3

Intro

du

ction

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system

poses a different set of proteolytic challenges We would

like to solicit feedback from researchers who have

encountered problems with standard cocktail formulations

as well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

IntroductionRobert GatesProduct Managerrobertgatessialcom

Proteolytic hydrolysis of peptide bonds was first studied as a function of digestion in higher mammals It is now recognized as an essential and ubiquitous mechanism for the regulation of a myriad of physiological processes

Inhibition of proteolytic activity is usually employed for two purposes

For the prevention of unwanted degradation of proteins during their isolation and characterization

To study the regulatory aspects of specific proteolytic events as they relate to cellular processes Some of the most studied proteolytic processes include blood coagulation and complement cascades hormonal regulation apoptosis extracellular matrix degradation proteasome and lysozomal regulation and disease states such as Alzheimerrsquos and viral replication

Four main classes of proteolyic enzymes have been routinely utilized to describe proteases The serine proteases are probably the best characterized This class of proteases includes trypsin chymotrypsin and elastase The cysteine protease class includes papain calpain and lysozomal cathepsins Aspartic proteases include pepsin and rennin Metalloproteinases include thermolysin and carboxypeptidase A

During isolation and characterization one or all four classes of proteases may pose a threat to the fate of a protein Broad-spectrum protease inhibitors and mixtures (or cocktails) have been developed to protect the integrity of isolated proteins Sigmareg offers and manufactures the broadest range of protease inhibitors and inhibitor cocktails of any supplier Sigma inhibitor cocktails have been specifically formulated for particular applications as they relate to the biological source or method of expression

Protease inhibitors can be added during cell growth and protein expression or can be added at the time of extraction

4 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Bro

ad S

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tru

m In

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ito

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f Pr

ote

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Enzy

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Cla

sses

Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics

Typical Working Concentrations

Preparation of Stock Solutions

Serine Protease Inhibitors

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM 100 mM in water stable 1 month at -20 degC

A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC

D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC

T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl

T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC

D0879 DIFP Highly toxic cholinesterase inhibitor Broad spectrum serine protease inhibitor Hydrolyzes rapidly in aqueous solutions

01 mM Prepare in anhydrous isopropanol at 200 mM

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Cysteine Protease Inhibitors

E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

Aspartic Protease Inhibitors

P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Metalloproteinase Inhibitors

ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water

P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC

R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase

1-10 mM 1 mM in water stable 1 month at -20 degC

B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data

Our Innovation Your Research mdash Shaping the Future of Life Science 5

Protease In

hib

itor Pan

el

Protease Inhibitor PanelProtease Inhibitor Panel

Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases

Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility

As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture

Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor

INHIB1-1KT 1 kit

INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility

AEBSF A8456 25 mg -20 degC 01-1 mM 2397 50 mgmL (water)

6-Aminohexanoic acid A2504 25 g RT 5 mgmL 1312 25 mgmL (water)

Antipain A6191 5 mg -20 degC 1-100 μM 6047 50 mgmL (water)

Aprotinin A1153 5 mg 2-8 degC 10-800 nM 6512 10 mgmL (water)

Benzamidine HCl B6506 5 g 2-8 degC 05-4 mM 1566 50 mgmL (water)

Bestatin B8385 5 mg -20 degC 40 mM 3448 25 mgmL (water)

Chymostatin C7268 5 mg -20 degC 6-60 μgmL (10-100 μM) ~608 6 mgmL (DMSO)

E-64 E3132 5 mg 2-8 degC 10 μM 3574 20 mgmL (water)

EDTA disodium salt ED2SS 50 g RT 1 mM 3722 50 mgmL (water)

N-Ethylmaleimide E3876 5 g 2-8 degC 01-1 mM 1251 50 mgmL (ethanol)

Leupeptin L2884 5 mg -20 degC 1-0-100 μM 4756 50 mgmL (water)

Pepstatin P5318 5 mg 2-8 degC 05-10 mgmL 6859 1 mgmL (ethanol)

Phosphoramidon R7385 5 mg -20 degC 10 mM (water) 5435 for free acid 10 mgmL (water)

Trypsin inhibitor T9003 100 mg -20 degC 11 stoichiometric binding 20100 20100 10 mgmL (water)

Visit sigmacomproteasefinder

Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location

Simple to use

Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)

capable of the cleavage

sigma-aldrichcom

Protease Finder

6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r C

ock

tails

an

d T

able

ts

Protease Inhibitor Cocktails and TabletsYou need a cocktail

End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg

Protease Inhibitor CocktailsCat No P2714 S8820 P1860 P8465 P8340 P8849 P8215 P9599

General UseLyophilized

powderWater Soluble

General UseTablet

Water Soluble

Tissue CultureSolution in

DMSO

Bacterial CellsLyophilized

powderVial of DMSO

Mammalian Tissues

Solution in DMSO

Histidine-Tagged Proteins

Solution in DMSO

FungalYeast Cells

Solution in DMSO

Plant CellsSolution in

DMSO

AEBSF

Aprotinin

Bestatin

EDTA

E-64

Leupeptin

Pepstatin A

Phosphoramidon

110-Phenanthroline

SIGMAFASTtrade Protease Inhibitor Tablets

For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)

One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract

S8820-20TAB 20 tablets

Protease Inhibitor Cocktail

for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA

One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin

P2714-1BTL 1 bottle

for use in purification of Histidine-tagged proteins DMSO solution

Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon

One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml

P8849-1ML 1 mL

P8849-5ML 5 mL

Life Science Innovationsmdashnew and emerging technologies put forth in a fresh unique way that applies to your area of study

BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic

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Download your copy of Life Science Innovations at sigmacominnovations

Subscribe or download your copy of BioFiles at sigmacombiofiles

A Perfect Fit

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

itor C

ocktails an

d Tab

lets

for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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dex

Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

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ct L

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ng

s

Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

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Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

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of continually curated biological data

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all with a single simple click

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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 2: Protease Inhibition and Detection

2

Life Science

BioFilesVolume 4 Number 2

Your gateway to Biochemicals and Reagents for Life Science Research from Sigmareg

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Register today for upcoming issues and eBioFiles announcements and receive a free IUBMB-Sigma-Nicholson Metabolic Pathway Chart sigmacombiofiles

Life Science TVFeaturing the Non-Specific Protease Activity Assay

The Sigmareg Life Science TV Web site is home to instructional informational and protocol based videos With regular updates to the library you will find videos covering many topics across the life science disciplines Currently available videos include

bull ATP Synthase Animation

bull GenomePlexreg Tutorial

bull TargeTronreg Technical Animation

The latest video addition to Life Science TV is titledNon-Specific Protease Activity Assay

This video displays a detailed protocol for determining the activity of proteases The universal assay procedure is useful for experiments ranging from sample preparation to QC analysis

View all Sigma Life Science Videos at sigma-aldrichcomlsvideos

Table of Contents

Introduction 3

Broad Spectrum Inhibitors of Proteolytic Enzyme Classes 4

Protease Inhibitor Panel 5

INHIB-1 Protease Inhibitor Panel Component Details 5

Protease Inhibitor Cocktails and Tablets 6

Protease Inhibitor Specificity Index 8

Product Listings 12

Aprotinin 15

E64 Analogs 20

Leupeptin 23

Trypsin Inhibitors 26

Protease Detection 29

Fluorescent Protease Detection 29

Non-Specific Protease Substrates 30

Technical content Christina Smith BS Robert Gates MS

Our Innovation Your Research mdash Shaping the Future of Life Science 3

Intro

du

ction

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system

poses a different set of proteolytic challenges We would

like to solicit feedback from researchers who have

encountered problems with standard cocktail formulations

as well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

IntroductionRobert GatesProduct Managerrobertgatessialcom

Proteolytic hydrolysis of peptide bonds was first studied as a function of digestion in higher mammals It is now recognized as an essential and ubiquitous mechanism for the regulation of a myriad of physiological processes

Inhibition of proteolytic activity is usually employed for two purposes

For the prevention of unwanted degradation of proteins during their isolation and characterization

To study the regulatory aspects of specific proteolytic events as they relate to cellular processes Some of the most studied proteolytic processes include blood coagulation and complement cascades hormonal regulation apoptosis extracellular matrix degradation proteasome and lysozomal regulation and disease states such as Alzheimerrsquos and viral replication

Four main classes of proteolyic enzymes have been routinely utilized to describe proteases The serine proteases are probably the best characterized This class of proteases includes trypsin chymotrypsin and elastase The cysteine protease class includes papain calpain and lysozomal cathepsins Aspartic proteases include pepsin and rennin Metalloproteinases include thermolysin and carboxypeptidase A

During isolation and characterization one or all four classes of proteases may pose a threat to the fate of a protein Broad-spectrum protease inhibitors and mixtures (or cocktails) have been developed to protect the integrity of isolated proteins Sigmareg offers and manufactures the broadest range of protease inhibitors and inhibitor cocktails of any supplier Sigma inhibitor cocktails have been specifically formulated for particular applications as they relate to the biological source or method of expression

Protease inhibitors can be added during cell growth and protein expression or can be added at the time of extraction

4 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Bro

ad S

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tru

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tic

Enzy

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sses

Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics

Typical Working Concentrations

Preparation of Stock Solutions

Serine Protease Inhibitors

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM 100 mM in water stable 1 month at -20 degC

A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC

D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC

T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl

T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC

D0879 DIFP Highly toxic cholinesterase inhibitor Broad spectrum serine protease inhibitor Hydrolyzes rapidly in aqueous solutions

01 mM Prepare in anhydrous isopropanol at 200 mM

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Cysteine Protease Inhibitors

E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

Aspartic Protease Inhibitors

P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Metalloproteinase Inhibitors

ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water

P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC

R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase

1-10 mM 1 mM in water stable 1 month at -20 degC

B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data

Our Innovation Your Research mdash Shaping the Future of Life Science 5

Protease In

hib

itor Pan

el

Protease Inhibitor PanelProtease Inhibitor Panel

Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases

Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility

As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture

Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor

INHIB1-1KT 1 kit

INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility

AEBSF A8456 25 mg -20 degC 01-1 mM 2397 50 mgmL (water)

6-Aminohexanoic acid A2504 25 g RT 5 mgmL 1312 25 mgmL (water)

Antipain A6191 5 mg -20 degC 1-100 μM 6047 50 mgmL (water)

Aprotinin A1153 5 mg 2-8 degC 10-800 nM 6512 10 mgmL (water)

Benzamidine HCl B6506 5 g 2-8 degC 05-4 mM 1566 50 mgmL (water)

Bestatin B8385 5 mg -20 degC 40 mM 3448 25 mgmL (water)

Chymostatin C7268 5 mg -20 degC 6-60 μgmL (10-100 μM) ~608 6 mgmL (DMSO)

E-64 E3132 5 mg 2-8 degC 10 μM 3574 20 mgmL (water)

EDTA disodium salt ED2SS 50 g RT 1 mM 3722 50 mgmL (water)

N-Ethylmaleimide E3876 5 g 2-8 degC 01-1 mM 1251 50 mgmL (ethanol)

Leupeptin L2884 5 mg -20 degC 1-0-100 μM 4756 50 mgmL (water)

Pepstatin P5318 5 mg 2-8 degC 05-10 mgmL 6859 1 mgmL (ethanol)

Phosphoramidon R7385 5 mg -20 degC 10 mM (water) 5435 for free acid 10 mgmL (water)

Trypsin inhibitor T9003 100 mg -20 degC 11 stoichiometric binding 20100 20100 10 mgmL (water)

Visit sigmacomproteasefinder

Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location

Simple to use

Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)

capable of the cleavage

sigma-aldrichcom

Protease Finder

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teas

e In

hib

ito

r C

ock

tails

an

d T

able

ts

Protease Inhibitor Cocktails and TabletsYou need a cocktail

End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg

Protease Inhibitor CocktailsCat No P2714 S8820 P1860 P8465 P8340 P8849 P8215 P9599

General UseLyophilized

powderWater Soluble

General UseTablet

Water Soluble

Tissue CultureSolution in

DMSO

Bacterial CellsLyophilized

powderVial of DMSO

Mammalian Tissues

Solution in DMSO

Histidine-Tagged Proteins

Solution in DMSO

FungalYeast Cells

Solution in DMSO

Plant CellsSolution in

DMSO

AEBSF

Aprotinin

Bestatin

EDTA

E-64

Leupeptin

Pepstatin A

Phosphoramidon

110-Phenanthroline

SIGMAFASTtrade Protease Inhibitor Tablets

For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)

One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract

S8820-20TAB 20 tablets

Protease Inhibitor Cocktail

for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA

One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin

P2714-1BTL 1 bottle

for use in purification of Histidine-tagged proteins DMSO solution

Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon

One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml

P8849-1ML 1 mL

P8849-5ML 5 mL

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BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic

sigma-aldrichcom

Download your copy of Life Science Innovations at sigmacominnovations

Subscribe or download your copy of BioFiles at sigmacombiofiles

A Perfect Fit

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

itor C

ocktails an

d Tab

lets

for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

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2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

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Page 3: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 3

Intro

du

ction

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system

poses a different set of proteolytic challenges We would

like to solicit feedback from researchers who have

encountered problems with standard cocktail formulations

as well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

IntroductionRobert GatesProduct Managerrobertgatessialcom

Proteolytic hydrolysis of peptide bonds was first studied as a function of digestion in higher mammals It is now recognized as an essential and ubiquitous mechanism for the regulation of a myriad of physiological processes

Inhibition of proteolytic activity is usually employed for two purposes

For the prevention of unwanted degradation of proteins during their isolation and characterization

To study the regulatory aspects of specific proteolytic events as they relate to cellular processes Some of the most studied proteolytic processes include blood coagulation and complement cascades hormonal regulation apoptosis extracellular matrix degradation proteasome and lysozomal regulation and disease states such as Alzheimerrsquos and viral replication

Four main classes of proteolyic enzymes have been routinely utilized to describe proteases The serine proteases are probably the best characterized This class of proteases includes trypsin chymotrypsin and elastase The cysteine protease class includes papain calpain and lysozomal cathepsins Aspartic proteases include pepsin and rennin Metalloproteinases include thermolysin and carboxypeptidase A

During isolation and characterization one or all four classes of proteases may pose a threat to the fate of a protein Broad-spectrum protease inhibitors and mixtures (or cocktails) have been developed to protect the integrity of isolated proteins Sigmareg offers and manufactures the broadest range of protease inhibitors and inhibitor cocktails of any supplier Sigma inhibitor cocktails have been specifically formulated for particular applications as they relate to the biological source or method of expression

Protease inhibitors can be added during cell growth and protein expression or can be added at the time of extraction

4 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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ad S

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Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics

Typical Working Concentrations

Preparation of Stock Solutions

Serine Protease Inhibitors

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM 100 mM in water stable 1 month at -20 degC

A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC

D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC

T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl

T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC

D0879 DIFP Highly toxic cholinesterase inhibitor Broad spectrum serine protease inhibitor Hydrolyzes rapidly in aqueous solutions

01 mM Prepare in anhydrous isopropanol at 200 mM

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Cysteine Protease Inhibitors

E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

Aspartic Protease Inhibitors

P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Metalloproteinase Inhibitors

ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water

P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC

R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase

1-10 mM 1 mM in water stable 1 month at -20 degC

B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data

Our Innovation Your Research mdash Shaping the Future of Life Science 5

Protease In

hib

itor Pan

el

Protease Inhibitor PanelProtease Inhibitor Panel

Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases

Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility

As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture

Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor

INHIB1-1KT 1 kit

INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility

AEBSF A8456 25 mg -20 degC 01-1 mM 2397 50 mgmL (water)

6-Aminohexanoic acid A2504 25 g RT 5 mgmL 1312 25 mgmL (water)

Antipain A6191 5 mg -20 degC 1-100 μM 6047 50 mgmL (water)

Aprotinin A1153 5 mg 2-8 degC 10-800 nM 6512 10 mgmL (water)

Benzamidine HCl B6506 5 g 2-8 degC 05-4 mM 1566 50 mgmL (water)

Bestatin B8385 5 mg -20 degC 40 mM 3448 25 mgmL (water)

Chymostatin C7268 5 mg -20 degC 6-60 μgmL (10-100 μM) ~608 6 mgmL (DMSO)

E-64 E3132 5 mg 2-8 degC 10 μM 3574 20 mgmL (water)

EDTA disodium salt ED2SS 50 g RT 1 mM 3722 50 mgmL (water)

N-Ethylmaleimide E3876 5 g 2-8 degC 01-1 mM 1251 50 mgmL (ethanol)

Leupeptin L2884 5 mg -20 degC 1-0-100 μM 4756 50 mgmL (water)

Pepstatin P5318 5 mg 2-8 degC 05-10 mgmL 6859 1 mgmL (ethanol)

Phosphoramidon R7385 5 mg -20 degC 10 mM (water) 5435 for free acid 10 mgmL (water)

Trypsin inhibitor T9003 100 mg -20 degC 11 stoichiometric binding 20100 20100 10 mgmL (water)

Visit sigmacomproteasefinder

Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location

Simple to use

Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)

capable of the cleavage

sigma-aldrichcom

Protease Finder

6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

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Protease Inhibitor Cocktails and TabletsYou need a cocktail

End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg

Protease Inhibitor CocktailsCat No P2714 S8820 P1860 P8465 P8340 P8849 P8215 P9599

General UseLyophilized

powderWater Soluble

General UseTablet

Water Soluble

Tissue CultureSolution in

DMSO

Bacterial CellsLyophilized

powderVial of DMSO

Mammalian Tissues

Solution in DMSO

Histidine-Tagged Proteins

Solution in DMSO

FungalYeast Cells

Solution in DMSO

Plant CellsSolution in

DMSO

AEBSF

Aprotinin

Bestatin

EDTA

E-64

Leupeptin

Pepstatin A

Phosphoramidon

110-Phenanthroline

SIGMAFASTtrade Protease Inhibitor Tablets

For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)

One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract

S8820-20TAB 20 tablets

Protease Inhibitor Cocktail

for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA

One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin

P2714-1BTL 1 bottle

for use in purification of Histidine-tagged proteins DMSO solution

Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon

One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml

P8849-1ML 1 mL

P8849-5ML 5 mL

Life Science Innovationsmdashnew and emerging technologies put forth in a fresh unique way that applies to your area of study

BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic

sigma-aldrichcom

Download your copy of Life Science Innovations at sigmacominnovations

Subscribe or download your copy of BioFiles at sigmacombiofiles

A Perfect Fit

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

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for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

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2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

Your gateway to products services and more life science research

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Read current and previous issues and register to receive future Biofiles issues

Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

Prod

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

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Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 4: Protease Inhibition and Detection

4 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Broad Spectrum Inhibitors of Proteolytic Enzyme ClassesCat No Inhibitor Characteristics

Typical Working Concentrations

Preparation of Stock Solutions

Serine Protease Inhibitors

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

A8456 AEBSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM 100 mM in water stable 1 month at -20 degC

A1153 Aprotinin Does not inhibit thrombin or factor Xa 03 μM or equimolar Freely soluble in water and stable at 2-8 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A9141 Antithrombin III Inhibits thrombin kallikreins plasmin trypsin and factors Ixa Xa and Xia equimolar Soluble at 10 unmL in water prepare stock solutions at neutral pH store at -20 degC

D7910 34-Dichloroisocoumarin Broad spectrum serine protease inhibitor 5-100 μM 10 mM in DMSO stable at -20 degC

T7254 TLCK Inhibits trypsin-like serine proteases 10-100 μM Prepare fresh at 10 mM in 1 mM HCl

T4376 TPCK Inhibits chymotrypsin-like serine proteases 10-100 μM 10 mM in ethanol stable at 4 degC

D0879 DIFP Highly toxic cholinesterase inhibitor Broad spectrum serine protease inhibitor Hydrolyzes rapidly in aqueous solutions

01 mM Prepare in anhydrous isopropanol at 200 mM

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Cysteine Protease Inhibitors

E3876 N-Ethylmaleimide Binds stoichiometrically to SH groups equimolar Water soluble at gt10mgmL prepare fresh

L2884 Leupeptin Inhibits trypsin-like serine proteases such as trypsin chymotrypsin chymase pepsin and thrombin Inhibits selected cysteine proteases such as calpain cathepsin BH amp L and papain

10-100 μM 10 mM in water stable 6 months at -20 degC

E3132 E-64 Will not inhibit serine protease with the exception of trypsin 1-10 μM 1 mM in water stable at -20 degC

C7268 Chymostatin Inhibits chymotrypsin-like serine proteases such as chymase cathepsins ABD and G Also inhibits some cysteine proteases such as papain

10-100 μM 10 mM in DMSO stable at -20 degC

A6191 Antipain Inhibits serine proteases such as plasmin thrombin and trypsin Also inhibits some cysteine proteases such as calpain and papain

1-100 μM 10 mM in water stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

P7626 PMSF Broad spectrum serine protease inhibitor Also reported to inhibit some cysteine proteases such as papain

01-10 mM Prepare fresh in anhydrous ethanol or isopropanol at 200 mM

Aspartic Protease Inhibitors

P5318 Pepstatin A Inhibits aspartic proteases such as renin chymosin and pepsin 1 μM 1 mM in methanol or DMSO stable at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

Metalloproteinase Inhibitors

ED2SS EDTA Broad spectrum metalloproteinase inhibitor 1-10 mM Very soluble and stable in water

P9375 110-Phenanthroline Broad spectrum metalloproteinase inhibitor 1-10 mM 200mM in methanol or DMSO stable at -20 degC

R7385 Phosphoramidon Strong inhibitor of metalloendoproteinases thermolysin and elastases but a weak inhibitor of collagenase

1-10 mM 1 mM in water stable 1 month at -20 degC

B8385 Bestatin Inhibitor of aminopeptidases 1-10 μM 1 mM in methanol stable 1 month at -20 degC

M6159 α2-Macroglobulin Broad spectrum protease inhibitor equimolar water soluble stable at -20 degC

References1 Proteolytic Enzymes A Practical Approach R J Benyon and J S Bond Eds pp 241-249 (1994)2 Handbook of Enzyme Inhibitors 2nd ed H Zollner (1993)3 Sigma Data

Our Innovation Your Research mdash Shaping the Future of Life Science 5

Protease In

hib

itor Pan

el

Protease Inhibitor PanelProtease Inhibitor Panel

Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases

Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility

As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture

Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor

INHIB1-1KT 1 kit

INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility

AEBSF A8456 25 mg -20 degC 01-1 mM 2397 50 mgmL (water)

6-Aminohexanoic acid A2504 25 g RT 5 mgmL 1312 25 mgmL (water)

Antipain A6191 5 mg -20 degC 1-100 μM 6047 50 mgmL (water)

Aprotinin A1153 5 mg 2-8 degC 10-800 nM 6512 10 mgmL (water)

Benzamidine HCl B6506 5 g 2-8 degC 05-4 mM 1566 50 mgmL (water)

Bestatin B8385 5 mg -20 degC 40 mM 3448 25 mgmL (water)

Chymostatin C7268 5 mg -20 degC 6-60 μgmL (10-100 μM) ~608 6 mgmL (DMSO)

E-64 E3132 5 mg 2-8 degC 10 μM 3574 20 mgmL (water)

EDTA disodium salt ED2SS 50 g RT 1 mM 3722 50 mgmL (water)

N-Ethylmaleimide E3876 5 g 2-8 degC 01-1 mM 1251 50 mgmL (ethanol)

Leupeptin L2884 5 mg -20 degC 1-0-100 μM 4756 50 mgmL (water)

Pepstatin P5318 5 mg 2-8 degC 05-10 mgmL 6859 1 mgmL (ethanol)

Phosphoramidon R7385 5 mg -20 degC 10 mM (water) 5435 for free acid 10 mgmL (water)

Trypsin inhibitor T9003 100 mg -20 degC 11 stoichiometric binding 20100 20100 10 mgmL (water)

Visit sigmacomproteasefinder

Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location

Simple to use

Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)

capable of the cleavage

sigma-aldrichcom

Protease Finder

6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r C

ock

tails

an

d T

able

ts

Protease Inhibitor Cocktails and TabletsYou need a cocktail

End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg

Protease Inhibitor CocktailsCat No P2714 S8820 P1860 P8465 P8340 P8849 P8215 P9599

General UseLyophilized

powderWater Soluble

General UseTablet

Water Soluble

Tissue CultureSolution in

DMSO

Bacterial CellsLyophilized

powderVial of DMSO

Mammalian Tissues

Solution in DMSO

Histidine-Tagged Proteins

Solution in DMSO

FungalYeast Cells

Solution in DMSO

Plant CellsSolution in

DMSO

AEBSF

Aprotinin

Bestatin

EDTA

E-64

Leupeptin

Pepstatin A

Phosphoramidon

110-Phenanthroline

SIGMAFASTtrade Protease Inhibitor Tablets

For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)

One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract

S8820-20TAB 20 tablets

Protease Inhibitor Cocktail

for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA

One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin

P2714-1BTL 1 bottle

for use in purification of Histidine-tagged proteins DMSO solution

Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon

One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml

P8849-1ML 1 mL

P8849-5ML 5 mL

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BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic

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A Perfect Fit

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

itor C

ocktails an

d Tab

lets

for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

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2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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uct Listin

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

gs

Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e D

etec

tio

n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Internet sigma-aldrichcom

Page 5: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 5

Protease In

hib

itor Pan

el

Protease Inhibitor PanelProtease Inhibitor Panel

Create your own broad-spectrum protease inhibitor cocktails or screen your extracts for proteolytic activity Panel includes inhibitors for serine cysteine calpains and metalloproteinases

Stock solutions of each inhibitor should be prepared first prior to creating a cocktail Mixtures of some inhibitor stock solutions may result in precipitation due to interactions between inhibitors and the mixing of solvents In most cases further dilution will aid solubility

As powders all reagents can be stored at 0 degC Those reagents designated for room temperature and 2-8 degC storage do not require storage at 0 degC but will not be adversely affected when stored at 0 degC Allow all powders to warm to room temperature before opening Store tightly sealed and protect from moisture

Panel components also include economical alternatives such as NEM EACA EDTA and soybean trypsin inhibitor

INHIB1-1KT 1 kit

INHIB-1 Protease Inhibitor Panel Component DetailsProtease Inhibitor Cat No Package Size Storage Temp Working Range Molecular Weight Stock Solution Solubility

AEBSF A8456 25 mg -20 degC 01-1 mM 2397 50 mgmL (water)

6-Aminohexanoic acid A2504 25 g RT 5 mgmL 1312 25 mgmL (water)

Antipain A6191 5 mg -20 degC 1-100 μM 6047 50 mgmL (water)

Aprotinin A1153 5 mg 2-8 degC 10-800 nM 6512 10 mgmL (water)

Benzamidine HCl B6506 5 g 2-8 degC 05-4 mM 1566 50 mgmL (water)

Bestatin B8385 5 mg -20 degC 40 mM 3448 25 mgmL (water)

Chymostatin C7268 5 mg -20 degC 6-60 μgmL (10-100 μM) ~608 6 mgmL (DMSO)

E-64 E3132 5 mg 2-8 degC 10 μM 3574 20 mgmL (water)

EDTA disodium salt ED2SS 50 g RT 1 mM 3722 50 mgmL (water)

N-Ethylmaleimide E3876 5 g 2-8 degC 01-1 mM 1251 50 mgmL (ethanol)

Leupeptin L2884 5 mg -20 degC 1-0-100 μM 4756 50 mgmL (water)

Pepstatin P5318 5 mg 2-8 degC 05-10 mgmL 6859 1 mgmL (ethanol)

Phosphoramidon R7385 5 mg -20 degC 10 mM (water) 5435 for free acid 10 mgmL (water)

Trypsin inhibitor T9003 100 mg -20 degC 11 stoichiometric binding 20100 20100 10 mgmL (water)

Visit sigmacomproteasefinder

Cleave proteins exactly where you want with Sigmarsquos Protease Finder The Protease Finder will identify the protease needed to cleave a specific peptide sequence at your desired location

Simple to use

Select either Endo- or Exoproteolytic cleavage Enter your protein sequence into the positional boxes Submit the request to instantly receive the protease(s)

capable of the cleavage

sigma-aldrichcom

Protease Finder

6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r C

ock

tails

an

d T

able

ts

Protease Inhibitor Cocktails and TabletsYou need a cocktail

End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg

Protease Inhibitor CocktailsCat No P2714 S8820 P1860 P8465 P8340 P8849 P8215 P9599

General UseLyophilized

powderWater Soluble

General UseTablet

Water Soluble

Tissue CultureSolution in

DMSO

Bacterial CellsLyophilized

powderVial of DMSO

Mammalian Tissues

Solution in DMSO

Histidine-Tagged Proteins

Solution in DMSO

FungalYeast Cells

Solution in DMSO

Plant CellsSolution in

DMSO

AEBSF

Aprotinin

Bestatin

EDTA

E-64

Leupeptin

Pepstatin A

Phosphoramidon

110-Phenanthroline

SIGMAFASTtrade Protease Inhibitor Tablets

For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)

One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract

S8820-20TAB 20 tablets

Protease Inhibitor Cocktail

for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA

One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin

P2714-1BTL 1 bottle

for use in purification of Histidine-tagged proteins DMSO solution

Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon

One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml

P8849-1ML 1 mL

P8849-5ML 5 mL

Life Science Innovationsmdashnew and emerging technologies put forth in a fresh unique way that applies to your area of study

BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic

sigma-aldrichcom

Download your copy of Life Science Innovations at sigmacominnovations

Subscribe or download your copy of BioFiles at sigmacombiofiles

A Perfect Fit

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

itor C

ocktails an

d Tab

lets

for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money

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Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

Prod

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

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Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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s

Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

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Imagine a tool that gives you the power

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 6: Protease Inhibition and Detection

6 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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ito

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ts

Protease Inhibitor Cocktails and TabletsYou need a cocktail

End your worries and start relaxing with Sigmareg protease inhibitor cocktails Everything you need for effective protease inhibition is included in one bottle Carefully manufactured with unvaried composition our cocktails allow you to protect your target proteins with confidence while gaining the efficiency of purchasing one product through one source Sigma-Aldrichreg

Protease Inhibitor CocktailsCat No P2714 S8820 P1860 P8465 P8340 P8849 P8215 P9599

General UseLyophilized

powderWater Soluble

General UseTablet

Water Soluble

Tissue CultureSolution in

DMSO

Bacterial CellsLyophilized

powderVial of DMSO

Mammalian Tissues

Solution in DMSO

Histidine-Tagged Proteins

Solution in DMSO

FungalYeast Cells

Solution in DMSO

Plant CellsSolution in

DMSO

AEBSF

Aprotinin

Bestatin

EDTA

E-64

Leupeptin

Pepstatin A

Phosphoramidon

110-Phenanthroline

SIGMAFASTtrade Protease Inhibitor Tablets

For General UseTableted formulation containing water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and EDTA (sodium salt)

One tablet makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin One tablet is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract

S8820-20TAB 20 tablets

Protease Inhibitor Cocktail

for general use lyophilized powderMixture of water-soluble protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) E-64 bestatin leupeptin aprotinin and sodium EDTA

One bottle makes 100 mL of cocktail One mL is recommended for the inhibition of proteases equivalent to 1 mg of USP pancreatin

P2714-1BTL 1 bottle

for use in purification of Histidine-tagged proteins DMSO solution

Mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases and thermolysin-like activities Formulated with no metal chelators that might inhibit binding of histidine-tagged proteins to metal affinity resins (IMAC) Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatin A E-64 and phosphoramidon

One mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli or 10 g of baculovirus-infected Spodoptera frugiperda pupal ovary cells in a total volume of 100 ml

P8849-1ML 1 mL

P8849-5ML 5 mL

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BioFilesmdashtailored for the life science researcher aligns our vast array of products within a relevant research topic

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Download your copy of Life Science Innovations at sigmacominnovations

Subscribe or download your copy of BioFiles at sigmacombiofiles

A Perfect Fit

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

itor C

ocktails an

d Tab

lets

for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

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2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

Your gateway to products services and more life science research

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Read current and previous issues and register to receive future Biofiles issues

Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money

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Received your first research grantFocus on your discoveries ndash not your expenses Let Sigma-Aldrich help you make the most of your research dollars

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

sigma-aldrichcom

24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Internet sigma-aldrichcom

Page 7: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 7

Protease In

hib

itor C

ocktails an

d Tab

lets

for use with mammalian cell and tissue extracts DMSO solutionA mixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatinA E-64 bestatin leupeptin and aprotinin Contains no metal chelators

One mL is recommended for the inhibition of proteases extracted from 20 g of bovine liver

P8340-1ML 1 mL

P8340-5ML 5 mL

for use in tissue culture media DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and aminopeptidases Contains aprotinin bestatin leupeptin E-64 and pepstatin A Contains no metal chelators

Use at a dilution of 1200 or more in tissue culture media to prevent proteolytic degradation of secreted proteins

Solution in DMSO (D 2650 Hybri-Max)

P1860-1ML 1 mL

for use with bacterial cell extracts lyophilized powderMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 bestatin and sodium EDTA

Five mL is recommended for the inhibition of proteases extracted from 20 g of Escherichia coli

Supplied with a vial of DMSO

P8465-5ML 5 mL

P8465-25ML 25 mL

for use with fungal and yeast extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metallo-proteases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) pepstatin A E-64 and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 20 g of yeast

P8215-1ML 1 mL

P8215-5ML 5 mL

for plant cell and tissue extracts DMSO solutionMixture of protease inhibitors with broad specificity for the inhibition of serine cysteine aspartic and metalloproteases and aminopeptidases Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF) bestatin pepstatinA E-64 leupeptin and 110-phenanthroline

One mL is recommended for the inhibition of proteases extracted from 30 g of plant tissue in a total volume of 100 ml

Tested for inhibition in extracts from Phaseolus vulgaris (kidney bean) Pisum sativum (pea) Triticum aestivum (wheat) Nicotiana tobaccum (tobacco) and Arabidopsis thaliana (arabidopsis)

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

No

Act

ivity

No

Act

ivity

12

1

08

06

04

02

0

Source

Aminopeptidase activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Am

ino

pep

tid

ase

acti

vity

(ug

tis

sue)

1

08

06

04

02

0

Source

Serine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

Seri

ne

pro

teas

e ac

tivi

ty(u

g t

issu

e)

03

025

02

015

01

005

0

Asp

arti

c p

rote

ase

acti

vity

(ug

tis

sue)

Source

Aspartic protease activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

00140012001

0008000600040002

0

Ch

ymo

tryp

sin

-lik

e ac

tivi

ty(u

g t

issu

e)

Source

Chymotrypsin-like activity

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

No

Act

ivity

218161412

108060402

0

Cys

tein

e p

rote

ase

acti

vity

(ug

tis

sue)

Source

Cysteine protease activity

as iswith inhibitor cocktail

beansprouts

peasprouts

pea leaves

pearoots

wheat sprouts

wheat leaves

wheat roots

tobacco sprouts

tobacco leaves

arabidopsis sprouts

Endogenous protease activities in plant tissues and their inhibition by Sigma-Aldrich plant cell protease inhibitor cocktail

P9599-1ML 1 mL

P9599-5ML 5 mL

While Sigma scientists have invested

considerable resources to formulate

the most effective cocktails suitable

for as many applications as possible

each cell line extractionpurification

procedure and expression system poses a different

set of proteolytic challenges We would like to solicit

feedback from researchers who have encountered

problems with standard cocktail formulations as

well as those who have developed novel alternative

strategies for protease inhibition Visit our BioBlog at

sigmabioblogscom to learn more

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Read current and previous issues and register to receive future Biofiles issues

Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

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Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 8: Protease Inhibition and Detection

8 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Cat No

Aminopeptidase

Bestatin B8385

Cysteamine M6500

Angiotensin Converting Enzyme

Angiotensin Converting Enzyme Inhibitor A0773

Bradykinin potentiator B B0507

Bradykinin potentiator C B0632

Captopril C4042

Captopril C8856

Fenoldopam F6800

Foroxymithine F5017

N-Succinyl-l-proline S6633

Bromelain

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Calpain

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

4-Chloromercuribenzoic acid C5913

E-64c E0514

E-64d E8640

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

N-Methoxysuccinyl-Val-Homo-Phe-fluoromethyl ketone M5439

Z-Leu-Leu-Leu-fluoromethyl ketone C8984

Calpain I

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

Calpain II

Acetyl-Calpastatin (184-210) human C4285

Antipain A6191

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Calpain Inhibitor Peptide C9181

4-Chloromercuribenzoic acid C5913

E-64c E0514

N-Ethylmaleimide E3876

Leupeptin L2023

α2-Macroglobulin M6159

Carboxypeptidase A

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Product Cat No

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase B

Carboxypeptidase Inhibitor C0279

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Carboxypeptidase P

4-Chloromercuribenzoic acid C5913

Diethyl pyrocarbonate D5758

Diisopropylfluorophosphate D0879

Sodium iodoacetate I2512

Carboxypeptidase Y

Aprotinin A1153

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Caspase

Boc-Asp(OMe)-fluoromethyl ketone B2682

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C8484

Caspase 1

N-Acetyl-Trp-Glu-His-Asp-al A1466

N-Acetyl-Tyr-Val-Ala-Asp chloromethyl ketone A4211

N-Acetyl-Tyr-Val-Lys(biotinyl)-Asp 26- dimethylbenzoyloxymethyl Ketone

A4339

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone C9484

Caspase 2

N-Acetyl-Val-Asp-Val-Ala-Asp-CHO A2222

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoromethyl ketone C1605

Caspase 3

N-Acetyl-Asp-Glu-Val-Asp-al A0835

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-VAD-FMK V116

Caspase 5

N-Acetyl-Trp-Glu-His-Asp-al A1466

Caspase 6

N-Acetyl-Val-Glu-Ile-Asp-al A6339

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoromethyl ketone C0480

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoromethyl ketone C1730

Caspase 7

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone C0605

Caspase 8

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone C1230

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone C8734

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

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2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

Starting a new labLet Sigma-Aldrichreg help you get it done on time on the spot and on the money

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

Prod

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

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Visit sigmacomenzymeexplorer

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Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 9: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 9

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

Caspase 9

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoromethyl ketone C1355

Caspase 13

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoromethyl ketone C8859

Cathepsin B

Antipain A6191

CA-074 C5732

CA-074 methyl ester C5857

Calpain Inhibitor I A6185

Calpain Inhibitor II A6060

Chymostatin C7268

Cystatin C0408

Cystatin C8917

E-64 E3132

Leupeptin L2023

α2-Macroglobulin M6159

Procathepsin B Fragment 26-50 rat P0350

Procathepsin B Fragment 36-50 rat P0475

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin C

Sodium iodoacetate I2512

Cathepsin D

Antipain A6191

Chymostatin C7268

Pepstatin A P5318

Cathepsin L

Calpain Inhibitor I A6185

Z-Phe-Phe-fluoromethyl ketone C9109

Cathepsin G

N-Acetyl-eglin C E7888

α1-Antichymotrypsin A9285

Chymostatin C7268

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Z-Gly-Leu-Phe-chloromethyl ketone C9984

α-Chymotrypsin

N-Acetyl-eglin C E7888

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

α1-Antichymotrypsin A9285

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bromoenol lactone B1552

Chymostatin C7268

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Product Cat No

Leupeptin L2023

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Trypsin-chymotrypsin inhibitor T9777

Trypsin inhibitor T0256

Trypsin inhibitor T9253

Trypsin inhibitor T9378

Trypsin inhibitor T4385

Clostripain

4-Chloromercuribenzoic acid C5913

Histatin 5 H6027

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Collagenase

Dichloromethylenediphosphonic acid D4434

Ethylenediaminetetraacetic acid E1644

Ethylenediaminetetraacetic acid ED2SS

Isoamylphosphonyl-Gly-Pro-Ala I7029

Complement C1

C1 Esterase Inhibitor E0518

Complement C1r

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement C1s

C1 Esterase Inhibitor E0518

Diisopropylfluorophosphate D0879

Complement Factor D

Diisopropylfluorophosphate D0879

Complement Factors B and D

Diisopropylfluorophosphate D0879

Dipeptidyl peptidase IV

Ile-Pro-Ile I9759

Dispase

Ethylenediaminetetraacetic acid ED2SS

110-Phenanthroline P9375

Elastase granulocyte

N-(Methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone M0398

Elastase leukocyte

N-Acetyl-eglin C E7888

Aprotinin A1153

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Elastase pancreatic

α1-Antitrypsin A9024

α1-Antitrypsin A6150

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Read current and previous issues and register to receive future Biofiles issues

Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

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LFL71112-508361

0039

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Page 10: Protease Inhibition and Detection

10 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e In

hib

ito

r Sp

ecif

icit

y In

dex

Product Cat No

Endoproteinase Arg-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Endoproteinase Glu-C

Diisopropylfluorophosphate D0879

α2-Macroglobulin M6159

Endoproteinase Lys-C

Aprotinin A1153

Diisopropylfluorophosphate D0879

Leupeptin L2023

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Factor VIIIa

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]

A0472

Factor Xa

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Trypsin inhibitor T9003

Factor XIa

C1 Esterase Inhibitor E0518

Factor XIIa

C1 Esterase Inhibitor E0518

Ficin

Cystatin C8917

E-64 E3132

N-Ethylmaleimide E3876

Sodium iodoacetate I2512

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Granzyme B

Antipain A6191

34-Dichloroisocoumarin D7910

Kallikrein Plasma

C1 Esterase Inhibitor E0518

Gabexate mesylate G2417

Kallikrein tissue

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Benzamidine B6506

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Leupeptin L2023

Product Cat No

Leucine Aminopeptidase

Actinonin A6671

Bestatin B8385

Leucine aminopeptidase cytosol

Actinonin A6671

Amastatin A1276

Bestatin B8385

34-Dichloroisocoumarin D7910

Diethylenetriaminepentaacetic acid D6518

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

110-Phenanthroline P9375

Leucine aminopeptidase microsomal

Actinonin A6671

Amastatin A1276

Bestatin B8385

Matrix metalloprotease

(R)-N4-Hydroxy-N1-[(S)-2-(1H-indol-3-yl)-1- methylcarbamoyl-ethyl]-2-isobutyl-succinamide

M5939

Metallo-protease inhibitor aminopeptidases M and B

l-Leucinethiol oxidized L8397

Metallo-protease inhibitor with selectivity for aminopeptidases

Epiamastatin E3389

Metallo-protease selectively aminopeptidases

Epibestatin E0381

Papain

Antipain A6191

Chymostatin C7268

Cystatin C8917

34-Dichloroisocoumarin D7910

E-64 E3132

Ebselen E3520

Gly-Gly-Tyr-Arg G5386

Leupeptin L2023

α2-Macroglobulin M6159

Nα-Tosyl-l-lysine chloromethyl ketone T7254

N-p-Tosyl-l-phenylalanine chloromethyl ketone T4376

Pepsin

Pepstatin A P5318

Plasmin

6-Aminocaproic acid A2504

6-Aminocaproic acid A7824

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Aprotinin A1153

Bdellin B3906

Protease Inhibitor Specificity IndexSee pages 12ndash30 for more details on each protease inhibitor

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

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Your gateway to products services and more life science research

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Join the Sigma-Aldrich New Lab Start-Up program ndash contact your local sales representative for more information

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

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gs

Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

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Features over 600 detailed procedures for measuring enzyme activity and related metabolites

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

gs

Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e D

etec

tio

n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Internet sigma-aldrichcom

Page 11: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 11

Protease In

hib

itor Sp

ecificity Ind

ex

Product Cat No

C1 Esterase Inhibitor E0518

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Elastatinal E0881

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethanesulfonyl fluoride P7626

Urinary Trypsin Inhibitor Fragment U4751

Pro-caspase 3

N-Acetyl-Glu-Ser-Met-Asp-al A1341

N-Acetyl-Ile-Glu-Thr-Asp-al A1216

Pronase E

Diisopropylfluorophosphate D0879

Ethylenediaminetetraacetic acid ED2SS

Protease HIV-1ANT

Acetyl-pepstatin A4815

Protease (Non-selective serine and cysteine protease inhibitor)

α-Boc-Deacetylleupeptin B7530

Protease of Bacteroides gingivalis

Histatin 5 H6027

Proteasome inhibitor

clasto-Lactacystin β-lactone L7035

Lactacystin L6785

Z-Leu-Leu-Phe-CHO C0358

Proteinase K

Diisopropylfluorophosphate D0879

Phenylmethanesulfonyl fluoride P7626

Renin

Pepstatin A P5318

β-Secretase

Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27- dimethyloctanoyl]-Ala-Glu-Phe

G8291

β-Secretase inhibitor S4562

γ-Secretase

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenylhexanoyl]-Leu-Phe-NH2

B5306

γ-Secretase inhibitor S2188

Thermolysin

Diethylenetriaminepentaacetic acid D6518

Diethylenetriaminepentaacetic acid D1133

Ethylenediaminetetraacetic acid ED2SS

Ethylene glycol-bis(2-aminoethylether)-NNNrsquoNrsquo-tetraacetic Acid E4378

α2-Macroglobulin M6159

110-Phenanthroline P9375

Phosphoramidon R7385

Product Cat No

Thrombin

N-Acetyl-eglin C E7888

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III A9141

Antithrombin III A2221

Antithrombin III A6397

34-Dichloroisocoumarin D7910

Diisopropylfluorophosphate D0879

Gabexate mesylate G2417

Hirudin H7016

Hirudin H7380

[Lys47]Hirudin (HV2) leech H0393

Leupeptin L2023

Phenylmethanesulfonyl fluoride P7626

Nα-Tosyl-l-lysine chloromethyl ketone T7254

Trypsin

4-(2-Aminoethyl)benzenesulfonyl fluoride A8456

Antipain A6191

Antithrombin III Bovine A9141

Antithrombin III Human A2221

α1-Antitrypsin A9024

α1-Antitrypsin A6150

Aprotinin A1153

Bdellin B3916

α2-Antiplasmin A0914

p-Aminobenzamidine dihydrochloride A7148

α1-Antitrypsin A9024

Benzamidine Hydrochloride B6506

Diisopropyl fluorophosphate D0879

34-Dichloroisocoumarin D7910

Ecotin E1398

Gabexate mesylate G2417

Leupeptin L2023

α2-Macroglobulin M6159

Phenylmethylsulfonyl fluoride P7626

Trypsin Inhibitor Bovine Pancreas T0256

Trypsin Inhibitor Chicken Egg T2011

Trypsin Inhibitor Turkey Egg T4385

Nα-p-Tosyl-l-lysine chloromethyl ketone hydrochloride T7254

Trypsin Inhibitor Soybean T9008

Trypsin Inhibitor Soybean T9128

Trypsin Inhibitor Chicken Egg T9253

Trypsin Inhibitor Lima Bean T9378

Trypsin-chymotrypsin inhibitor T9777

Urokinase

Glu-Gly-Arg-chloromethyl ketone G2917

12 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

du

ct L

isti

ng

s

Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

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Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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uct Listin

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

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Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 12: Protease Inhibition and Detection

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Product Listings

Acetyl-Ala-Leu-Cys-Asp-Asp-Pro-Arg-Val-Asp-Arg-Trp-Tyr-Cys-Gln-Phe-Val-Glu-Gly-NH2 [Disulfide bridge 3-13]E76 C97H139N27O29S2 FW 221144Potent inhibitor of coagulation factor VIIIa

ge95 (HPLC)A0472-1MG 1 mg

N-Acetyl-Asp-Glu-Val-Asp-alAc-DEVD-CHO [169332-60-9] C20H30N4O11 FW 50247Reversible inhibitor of IL-1β converting enzyme (ICE) inhibits poly(ADP-ribose) polymerase cleavage by apopain (caspase 3)

ge95 powderA0835-1MG 1 mg

A0835-5MG 5 mg

Acetyl-Calpastatin (184-210) humanCS peptide Acetyl-Calpain inhibitor fragment 184-210 Ac-Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala-NH2 [79079-11-1] C142H230N36O44S FW 317763Inhibitor of calpain which induces an increase in secreted amyloid β-protein 1-42

~95 (HPLC) powderC4285-5MG 05 mg

N-Acetyl-Glu-Ser-Met-Asp-alAc-ESMD-CHO C19H30N4O10S FW 50653

ge90 (HPLC) powderBlocks the formation of the active p17 subunit of caspase 3 from its precursor and induces accumulation of the p20 precursor peptide

A1341-1MG 1 mg

N-Acetyl-Ile-Glu-Thr-Asp-al

Ac-IETD-CHOC21H34N4O10 FW 50252

NH

HN

NH

HN

H

O

H3C

O

O

O

O

CH3

O OH

CH3HO

O

OH

CH3

~99 (TLC) powderCaspase 8 inhibitor blocks the cleavage of the 32 kDa caspase 3 precursor into the p12 and p20 subunits thus blocking the formation of active caspase 3

A1216-1MG 1 mg

N-Acetyl-Trp-Glu-His-Asp-alAc-WEHD-CHO C28H33N7O9 FW 61160Very potent caspase 1 and 5 inhibitor

ge80 (HPLC) powderA1466-1MG 1 mg

Refer to Protease Inhibitor Specificity Index on pages 8ndash11 for related enzymes

N-Acetyl-Tyr-Val-Lys(bio tin yl)-Asp 26-dimethyl benzoyl oxy -methyl keto neAc-YVK(bio tin yl)D 26-dimethyl benzoyl oxy methyl keto ne C46H63N7O12S FW 93810Useful for affinity labeling the larger subunit of activated caspase 1 and caspase 1 related proteases

ge90 (HPLC) powderA4339-1MG 1 mg

A4339-5MG 5 mg

N-Acetyl-Val-Asp-Val-Ala-Asp-CHOAc-VDVAD-CHO Ac-Val-Asp-Val-Ala-Asp-CHO [194022-51-0] C22H33N5O11 FW 54352Caspase 2 Inhibitor

ge90 (HPLC) solidA2222-5MG 5 mg

N-Acetyl-Val-Glu-Ile-Asp-alAc-VEID-CHO C22H36N4O9 FW 50054Caspase 6 inhibitor

~99 (HPLC) powderA6339-1MG 1 mg

A6339-5MG 5 mg

Actinonin

3-[[1-[(2-(Hydroxy methyl)-1-pyrrolidin yl) carbo nyl]-2-methyl pro pyl]carba moyl]octa no-hydro xamic acid[13434-13-4] C19H35N3O5 FW 38550

N

HNO

O

O NH

OH

OH

CH3

CH3

H3C

Inhibitor of leucine aminopeptidase

A6671-10MG 10 mg

A6671-25MG 25 mg

Amastatin hydrochloride hydrate

(2S3R)-3-Amino-2-hydroxy-5-methyl - hexa noyl-Val-Val-Asp hydrochloride hydrate [100938-10-1] C21H38N4O8 middot HCl middot xH2O FW 51101 (Anh)

HNNH

NH

OO

OO

OHO

NH2

HOOH

HCl

H3C

CH3

H3CCH3

CH3

CH3

bull H2O

Amastatin is a slow tight-binding inhibitor of aminopeptidases It inhibits cytosolic leucine aminopeptidase (EC34111) microsomal aminopeptidase M (EC34112) and bacterial leucine aminopeptidase (EC341110) It is less effective against aminopeptidase A (EC 34117) the enzyme that converts Angiotensin II to Angiotensin III Effective concentration 1-10 μM

ge97 (HPLC)A1276-250UG 250 μg

A1276-5MG 05 mg

A1276-1MG 1 mg

A1276-5MG 5 mg

A1276-10MG 10 mg

A1276-25MG 25 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

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Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 13: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 13

Prod

uct Listin

gs

2-Amino benz oyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-Tyr(NO2)-AspAn thra niloyl-Ala-Phe-Ala-Phe-Asp-Val-Phe-3-nitro-Tyr-Asp-OH C62H71N11O18 FW 125829Fluorogenic substrate for Asp-specific proteases from Staphylococcus aureus Bacillus lichenformis and Streptomyces griseus

gt90 (HPLC)A9971-1MG 1 mg

6-Amino caproic acid

EACA 6-Amino hexanoic acid ε-Amino caproic acid [60-32-2] H2N(CH2)5CO2H FW 13117 H2N

OH

O

Lysine analog Promotes rapid dissociation of plasmin thereby inhibiting the activation of plasminogen and subsequent fibrinolysis12 Reported to inhibit plasminogen binding to activated platelets3 An early report indicated that it inhibits the activation of the first component of the complement system4 Binds and inactivates Carboxypeptidase B5

Lit cited 1 Humphries JE et al Fibrinogenolytic and fibrinolytic activity of cell-associated plasmin Arterioscler Thromb 13 48-55 (1993) 2 Krishnamurti C et al Inhibitory effects of lysine analogues on t-PA induced whole blood clot lysis Thromb Res 73 419-430 (1994) 3 Adelman B et al Plasminogen interactions with platelets in plasma Blood 72 1530-1535 (1988) 4 Soter NA et al Inhibition by ε-aminocaproic acid of the activation of the first component of the complement system J Immunol 114 928 (1975) 5 Dessaint JP et al Catheptic carboxypeptidase B as a major component in ldquoT-cell activating factorrdquo of macrophages J Immunopharmacol 1 399-414 (1979)

ge99 (titration) powderEACA is reported to inhibit chymotrypsin Factor VIIa lysine carbo xy peptidase plasmin and plasminogen activator

EACA is directly soluble in water at 25 mgml As an inhibitor of plasmin it has been utilized in the clotting buffer for fibrinogen assays This buffer is 10 mM potassium and sodium phosphate pH 64 with 020 g CaCl2 5 g 6-Aminohexanoic acid 1 g sodium azide and 9 g NaCl in 1 liter The buffer is stable indefinitely at room temperature

A2504-25G 25 g

A2504-100G 100 g

A2504-500G 500 g

A2504-1KG 1 kg

A2504-5KG 5 kg

SigmaUltra ge99Insoluble matter le01Phosphorus (P) le0005

A7824-25G 25 g

A7824-100G 100 g

A7824-500G 500 g

A7824-1KG 1 kg

4-(2-Amino ethyl)ben zene sul fonyl fluoride hydrochloride

AEBSF [30827-99-7] C8H10FNO2S middot HCl FW 23969 S OO

H2N

F

bull HCl

Irreversible serine protease inhibitor Inhibition constants are similar to those of PMSF and DFP AEBSF has been shown to inhibit trypsin chymotrypsin plasmin kallikrein and thrombin As an alternative to PMSF and DFP AEBSF offers lower toxicity improved solubility in water and improved stability in aqueous solutions AEBSF has been used in cell culture in concentrations of up to 025 mM

Recommended concentrations for use are 01-1 mM

ge97A8456-25MG 25 mg

A8456-100MG 100 mg

A8456-500MG 500 mg

Angiotensin Converting Enzyme InhibitorpGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro [35115-60-7] C53H76N14O12 FW 110126

ge95 (TLC)A0773-1MG 1 mg

A0773-5MG 5 mg

α1-Anti chymotrypsin from human plasma[141176-92-3]Glycoprotein is a specific inhibitor of chymotrypsin-like serine proteases

~95 (SDS-PAGE) lyophilized powderLyophilized powder containing Tris buffer salt and NaCl

Package size based on protein content

A9285-1MG 01 mg

Anti pain dihydrochloride from microbial source

N-(Nα-Carbo nyl-Arg-Val-Arg-al)-Phe [37682-72-7] C27H44N10O6 middot 2 HCl FW 67762

NH

H

O

NH

NH

H2NO

HNCH3

CH3

ONH

H2NNH

NH

O

HN

O OH

bull 2HCl

Isolated from a microbial source antipain hydrochloride is a reversible inhibitor of serinecysteine proteases and some trypsin-like serine proteases Its action resembles leupeptin however its plasmin inhibition is less and its cathepsin A inhibition is more than that observed with leupeptin

Concentrations for 50 inhibition (μgml)

papain 016

trypsin 026

cathepsin A 119

cathepsin B 059

cathepsin D 125

plasmin gt93

chymotrypsin and pepsin gt250

It also has been reported to inhibit calpain I (porcine) with Ki = 14 μM

Solubility testing at 50 mgml in water yields a clear to slightly hazy yellow solution It is reportedly soluble in methanol water and DMSO less soluble in ethanol butanol and propanol insoluble in benzene hexane and chloroform8 A stock solution in water or buffer is stable for about a month at -20 degC

Dilute solutions should be stored on ice and kept for only a day because of the terminal aldehyde which is subject to oxidation and racemization

Stock solutions in water or buffer stable for 1 week at 4 degC 1 month at minus20 degC

A6191-1MG 1 mg

A6191-5MG 5 mg

A6191-25MG 25 mg

A6191-100MG 100 mg

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

sigma-aldrichcom

Your gateway to products services and more life science research

Visit sigmacombiofiles

Read current and previous issues and register to receive future Biofiles issues

Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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uct Listin

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

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Page 14: Protease Inhibition and Detection

14 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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q Antithrombin III

Factor Xa inhibitor Heparin cofactor [90170-80-2]Serine protease inhibitor Inhibitor of thrombin and activated factor X (Xa) Effective concentration equimolar with proteinase

Anti thrombin III from bovine plasma

lyophilized powder activity 200-400 unitsmg protein (E1

280 = 65)When reconstituted with 10 mL of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A9141-10UN 10 units

A9141-50UN 50 units

Anti thrombin III from human plasma

lyophilized powder ge95 (SDS-PAGE)Lyophilized from 20 mM Tris pH 80

One unit is the activity present in 01 mL of normal human pooled plasma tested in the presence of 01 unit of heparin

A2221-25UG 25 μg

A2221-125UG 125 μg

Anti thrombin III from rat plasma

lyophilized powder activity gt200 unitsmg protein (E1

280 = 65)When reconstituted with 10 ml of deionized water solution will contain a minimum of the stated activity in 01 M sodium chloride and 002 M Tris pH 75

One unit is the activity present in 01 ml of normal human pooled plasma tested in the presence of 01 unit of heparin

Homogeneous by gel electrophoresis

A6397-10UN 10 units

Antithrombin III p

α1-Anti trypsin from human plasmaα1-Protein ase inhibitor [9041-92-3]Serine protease inhibitor inhibits trypsin chymotrypsin and pancreatic and granulocytic elastase and acrosin Effective concentration equimolar with proteinase

Chromatographically prepared and partially purified

salt-free lyophilized powder1-4 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 2-6 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A9024-5MG 5 mg

A9024-25MG 25 mg

A9024-100MG 100 mg

salt-free lyophilized powder5-10 mg will inhibit 10 mg of trypsin with activity of 10000 BAEE units per mg protein 5-10 mg will inhibit approx 10 mg of α-chymotrypsin with activity of 40-50 BTEE units per mg protein

A6150-25MG 25 mg

A6150-100MG 100 mg

A6150-500MG 500 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 15

Prod

uct Listin

gs

Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

sigma-aldrichcom

24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 15: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 15

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Aprotinin

10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Aprotinin(bovine)

Propeptide

Propeptide

Kunitz domain

Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (~01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

Aprotinin Protease Inhibition Specificity

Enzyme Ki

Acrosin Weak inhibition

Chymotrypsin Ki = 9 nM

Chymotrypsinogen (bovine) pH 80 Ki = 9 nM

Elastase (human leukocytes) pH 80 Ki = 35 μM

Kallikrein (pancreatic) pH 80 Ki = 10 nM

Kallikrein (plasma) Ki = 30 nM 100 nM

Kallikrein (tissue) Ki = 1 nM

Kallikrein (urine) Ki = 17 nM

Plasmin (porcine) pH 78 Ki = 40 nM

Plasminogen activator Ki = 8 μM 27 μM

Trypsin (bovine) pH 80 Ki = 006 pM

Trypsinogen (bovine) pH 80 Ki = 18 μM

Tryptase TL-2 16 Inhibition at 10 μM

Urokinase (human) pH 88 Ki = 80 μM

q Aprotinin

Aprotinin bovine

BioUltra recombinant expressed in Nicotiana activity ge5 TIUmg protein ge98 (SDS-PAGE)

Contains no animal-derived components or impurities and is manufactured by transient expression of the aprotinin message in RNA (+)-strand tobacco mosaic virus vectors propagated in non-transgenic Nicotiana plants This is a recombinant form of the native bovine-sequence aprotinin which is traditionally isolated from bovine lung by methods involving fractional precipitation gel filtration and ion exchange chromatography Unlike animal-derived aprotinin this product is isolated and purified from plant tissue by proprietary methods

65142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6103) under reducing conditions and stained with Coomassie blue

A6103-1MG 1 mg

A6103-25MG 25 mg

A6103-100MG 100 mg

Aprotinin (Bovine)

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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Our Innovation Your Research mdash Shaping the Future of Life Science 17

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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gs

Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 16: Protease Inhibition and Detection

16 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Aprotinin from bovine lungTrypsin inhibitor (basic) [9087-70-1] C284H432N84O79S7 FW 651144Aprotinin is a competitive serine protease inhibitor that inhibits trypsin chymotrypsin kallikrein and plasmin Aprotinin forms stable complexes with and blocks the active sites of enzymes Binding is reversible with most aprotinin-protease complexes dissociating at pH gt10 or lt3 Effective concentration equimolar with protease

Aprotinin is a protein consisting of 58 amino acids arranged in a single polypeptide chain that is crosslinked by three disulfide bridges

Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit)

1 TIU sim 1300 KIU

saline solution activity 3-7 TIUmg proteinSolution in 09 NaCl and 09 benzyl alcohol

Protein determined by biuret

A6279-5ML 5 mL

A6279-10ML 10 mL

A6279-5X10ML 5 times 10 mL

A6279-50ML 50 mL

lyophilized powder activity 3-8 TIUmg solidA1153-1MG 1 mg

A1153-5MG 5 mg

A1153-10MG 10 mg

A1153-25MG 25 mg

A1153-100MG 100 mg

A1153-250MG 250 mg

BioUltra activity ge4 TIUmg solid ge98 (SDS-PAGE) 8Aprotinin is freely soluble in water (gt10 mgml) and in aqueous buffers of low ionic strengths Dilute solutions are generally less stable than concentrated ones Solution stability is pH dependent a range of 1-12 can be tolerated Repeated freeze-thaw cycles should be avoided Due to its compact tertiary structure aprotinin is relatively stable against denatura tion due to high temperature pH extremes organic solvents or proteolytic degradation (only thermolysin has been found capable of degrading aprotinin after heating to 60-80 degC) The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices but the use of acetylated materials and concentrated salt solutions (=01 M NaCl in buffer) minimizes the problem Sterilization may be achieved by filtration through a 02 μm filter

A further purification of A1153 to yield a product with slightly higher activity and purity

One trypsin inhibitor unit (TIU) will decrease the activity of 2 trypsin units by 50 where one trypsin unit will hydrolyze 10 μmole of N-α-benzoyl-Dl-arginine p-nitroanilide (BAPNA) per min at pH 78 at 25 degC

65

142

20

2429

3645556697116

200

SDS-PAGE (4-20) of aprotinin (Cat No A6106) under reducing conditions and stained with Coomassie blue

A6106-1MG 1 mg

A6106-25MG 25 mg

A6106-100MG 100 mg

Aprotinin p

Bdellin from leechesPlasmin inhibitor [62494-89-7]

lyophilized powder activity gt01 IUmg protein (E280)Mixture of bdellins A and B

Lyophilized powder containing glycine

Package size based on protein content

One inhibitory unit (IU) will reduce the plasmin catalyzed hydrolysis of the chromogenic substrate D-Ile-Pro-Arg-pNA by 10 μmole per min at pH 83 at 25 degC

B3906-1MG 01 mg

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sigma-aldrichcom

Our Innovation Your Research mdash Shaping the Future of Life Science 17

Prod

uct Listin

gs

Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

sigma-aldrichcom

24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 17: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 17

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Benz ami dine hydrochloride hydrate

Ami dino ben zene hydrochloride Benz amidi nium chloride Ben zene carbox imid amide hydrochloride [206752-36-5] C6H5C(=NH)NH2 middot HCl middot xH2O FW 15661 (Anh)

NH

NH2

bull HCl bull H2O

Benzamidine HCl is a reversible inhibitor of trypsin trypsin-like enzymes and serine proteases A concentration of approximately 1 mM is used for general protease inhibition To inhibit proteases from yeast a range of 05 to 40 mM is used and it is for the most part interchangeable with pepstatin A

In addition to being a strong competitive inhibitor of trypsin benzamidine HCl has been also shown to be a strong competitive inhibitor of thrombin and plasmin It was also found to be as effective as aprotinin in the prevention of glucagon degradation in human plasma

Benzamidine HCl is soluble in water and alcohol Solubility testing in water at a concentration of 50 mgml yields a clear solution with heating Benzamidine HCl is sensitive to oxidation It is recommended to prepare solutions fresh each time in degassed water prior to use However frozen aliquots stored under inert gas to exclude air may be stable for a short time Insufficient information is available to assess the shelf-life of a frozen solution

ge98B6506-5G 5 g

B6506-25G 25 g

B6506-100G 100 g

[(2R4R5S)-2-Benzyl-5-(Boc-amino)-4-hydroxy-6-phenyl hexa noyl]-Leu-Phe-NH2

l-685458 C39H52N4O6 FW 67285Potent γ-secretase inhibitor containing a hydroxy ethylene dipeptide isostere that should serve as a transition analogue to direct the inhibitor to the active site of an aspartyl protease target

gt90 (TLC)B5306-1MG 1 mg

Bestatin hydrochloride

N-[(2S3R)-3-Amino-2-hydroxy-4-phenyl-butyryl]-l-leu cine hydrochloride [65391-42-6] C16H24N2O4 middot HCl FW 34483

HN

O O

OH

H2NOH

HCl

CH3

CH3

A metalloprotease inhibitor selective for aminopeptidase Bestatin is a competitive and specific inhibitor of leucine aminopeptidase aminopeptidase B and triamino peptidase It inhibits aminopeptidase B at 60 nM (using arginine-β-naphthylamide as substrate) and leucine aminopeptidase at 20 nM (leucine-β-naphthylamide as substrate) It showed no inhibition of aminopeptidase A trypsin chymotrypsin elastase papain pepsin or themolysin

ge98 (HPLC)Solubility testing in water at 25 mgml yields a clear solution Stock solutions at 1 mM are expected to be stable at least one month stored at -20 degC

B8385-5MG 05 mg

B8385-1MG 1 mg

B8385-5MG 5 mg

B8385-10MG 10 mg

B8385-25MG 25 mg

B8385-100MG 100 mg

Boc-Asp(OMe)-fluoro methyl keto ne

Boc-D-FMK [187389-53-3] C11H18NO5F FW 26326

OCH3

ONH

OF

Boc

ge90 (TLC) solidCaspase inhibitor designed as a methyl ester to facilitate cell permeability

B2682-3MG 3 mg

α-Boc-De acetyl leupeptinNα-t-Boc-Leu-Leu-Arg-al[81344-47-0] C23H44N6O5 FW 48463

ge97 (TLC)B7530-5MG 5 mg

Bradykinin potentiator BpGlu-Gly-Leu-Pro-Pro-Arg-Pro-Lys-Ile-Pro-Pro [30892-86-5] C56H91N15O13 FW 118241

ge97 (TLC)B0507-5MG 5 mg

Bromo enol lactone

BEL E-6-(Bromo ethylene)tetra hydro-3-(1-naph thyl)-2H-pyran-2-one [88070-98-8] C16H13BrO2 FW 31718 O

O

Br H

Potent irreversible inhibitor of calcium-independent phospholipase A2 and of magnesium-dependent phosphatidate phosphohydrolase from P388D macrophages (IC50 = 8 μM) enzyme activated irreversible chymotrypsin inhibitor (Ki = 636 nM)

ge98B1552-5MG 5 mg

C1 Ester ase Inhibitor from human plasmaComplement C1 ester ase inhibitor Ester ase inhibitor C-1 from human plasma[80295-38-1]Esterase inhibitor C1 is a multifunctional regulator of all major kinin-generating protein cascade systems It has been a therapeutic tool in the treatment of hereditary angioedema and complement-mediated inflammatory tissue damage such as capillary leak syndrome septic shock multiple organ failure and hyperacute graft rejection

aqueous solution ge95 (SDS-PAGE)Solution in 20 mM potassium phosphate pH 70 containing 250 mM KCl

E0518-1MG 1 mg

CA-074

(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline [134448-10-5] C18H29N3O6 FW 38344

OHN

O

H

HNH

NO

O

H3C CH2CH3

COOHH3C

99 (TLC) solidInhibitor of Cathepsin B

C5732-5MG 5 mg

CA-074 methyl ester(L-3-trans-(Pro pyl carb amyl)oxirane-2-carbo nyl)-l-i sole ucyl-l-proline methyl ester [147859-80-1] C19H31N3O6 FW 39747

gt99 (HPLC) solidCA-074 methyl ester is an inhibitor of cathepsin B

C5857-5MG 5 mg

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

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Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 18: Protease Inhibition and Detection

18 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Calpain Inhibitor I

ALLN MG-101 Ac-LLnL-CHO N-Acetyl-Leu-Leu-Norleu-al N-Acetyl-l-leucyl-l-leucyl-l-norleucinal [110044-82-1] C20H37N3O4 FW 38353 H3C N

H

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

CH3

ge97 (TLC) powderCan initiate apoptosis in HL-60 cells1 but blocks dexamethasone-induced apoptosis in thymocytes and cycloheximide-induced apoptosis in metamyelocytes2 Inhibits cyclin B degradation and arrests the cell cycle at G1S and at meta-anaphase3 Blocks induction of nitric oxide synthase by LPS in macrophages4

Lit cited 1 Lu Q Mellgren RL Calpain inhibitors and serine protease inhibitors can produce apoptosis in HL-60 cells Arch Biochem Biophys 334 175 (1996) 2 Squier MK Calpain activation in apoptosis J Cell Physiol 159 229 (1994) 3 Griscavage JM Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-κ B Proc Natl Acad Sci USA 93 3308 (1996) 4 Sherwood SW In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetyl leucylleucyl norleucinal Proc Natl Acad Sci USA 90 3353 (1993)

A6185-5MG 5 mg

A6185-25MG 25 mg

Calpain Inhibitor II

N-Acetyl-l-leucyl-l-leucyl-l-me thio ninal [136632-32-1] C19H35N3O4S FW 40156

H3C NH

HN

NH

HO

O

O

O

CH3

CH3

CH3

CH3

SCH3

ge97 (TLC) powderA6060-5MG 5 mg

A6060-25MG 25 mg

Calpain Inhibitor PeptideAsp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-Leu-Ala [128578-18-7] C140H227N35O44S FW 313657Strong inhibitor of Calpain I and II but not of papain or trypsin

ge95 (HPLC)C9181-5MG 05 mg

C9181-1MG 1 mg

Captopril

N-[(S)-3-Mer capto-2-methyl pro pionyl]-l-proline [62571-86-2] C9H15NO3S FW 21729 N

O

OH

OHS

CH3

Angiotensin converting enzyme inhibitor Inhibits the formation of angiotensin II a bioactive peptide that stimulates angiogenesis and increases microvessel density

ge98 (HPLC) powderC4042-5G 5 g

C4042-25G 25 g

meets USP testing specificationsC8856-1G 1 g

C8856-5G 5 g

C8856-25G 25 g

Carboxy pepti dase Inhibitor from potato tuber[56092-22-9]

lyophilized powderOne μg of inhibitor protein will inhibit the activity of a minimum of 9 μg of carboxypeptidase A by 50 using hippuryl-l-phenylalanine as substrate at pH 75 at 25 degC

Lyophilized powder containing traces of ammonium bicarbonate buffer salts balance primarily non-protein components

Package size based on protein content

C0279-5MG 5 mg

C0279-10MG 10 mg

C0279-25MG 25 mg

4-Chloro mer curi benzoic acid

4-(Hydroxy mer curi)benzoic acid4-(Chloro mer curio)benzoic acid [59-85-8] ClHgC6H4CO2H FW 35716

OH

O

ClHg

C5913-5G 5 g

Chymostatin

N-(Nα-Carbo nyl-Cpd-X-Phe-al)-Phe (Cpd = capreomyci dine) (capreomyci dine = [SS]-α-(2-Imino hexa hydro-4-pyrimi dyl)gly cine) [9076-44-2]

HOOC CH NHCH2

CO

NH CH

NH

HN NH

CO

X Phenylalaninal

Chymostatin A X = Leu

Chymostatin B X = Val

Chymostatin C X = Ile

Chymostatin is a strong inhibitor of many proteases including chymotrypsin papain chymotrypsin-like serine proteinases chymases and lysosomal cysteine proteinases such as cathepsins ABC D H and L It weakly inhibits human leucocyte elastase It is effective at a final concentration of 100 to 200 μgml (10 to 100 μM) Chymostatin is often included in protease inhibitor cocktails used with plant extracts

Solubility testing in glacial acetic acid at 10 mgml yields a clear solution which is usually colorless but can be yellow in appearance It is reportedly also soluble in DMSO only slightly soluble in water and short-chain alcohols insoluble in ethyl acetate butyl acetate ether hexane and petroleum ether Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 degC Stock solutions can also be made in 01 μM HCl Dilute solutions (10-100 μM) are only stable for several hours due to oxidation of the terminal aldehyde

A mixture of A (major) B and C components A X=Leu B X=Val C X=Ile

chymostatin A MW = 6077

chymostatin B MW = 5937

chymostatin C MW = 6077

C7268-1MG 1 mg

C7268-5MG 5 mg

C7268-25MG 25 mg

C7268-50MG 50 mg

C7268-100MG 100 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 19

Prod

uct Listin

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clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 19: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 19

Prod

uct Listin

gs

clasto-Lactacystin β-lactone

[154226-60-5] C10H15NO4 FW 21323

NH

O

OH3C

H

OOH

CH3

CH3

Cell-permeable and irreversible proteasome inhibitor Lactacystin acts as a precursor for clasto-lactacystin β-lactone

L7035-1MG 01 mg

Cysta tin from chicken egg whiteCys teine protease inhibitor [81989-95-9]Cysteine proteases inhibitor including dipeptidyl peptidase III Effective concentration equimolar with protease

buffered aqueous glycerol solutionSolution in 10 mM Tris buffer pH 80 containing 50 glycerol

Protein determined by E1280

C0408-250UG 250 μg

C0408-5MG 05 mg

lyophilized powderPackage size based on protein content

C8917-5MG 05 mg

C8917-1MG 1 mg

Cyste amine hydrochloride

β-Mer capto ethyl amine hydrochloride Thio ethanol amine hydrochloride 2-Amino ethane thiol hydrochloride 2-Mer capto ethyl amine hydrochloride Deca rboxy cys teine hydrochloride [156-57-0] HSCH2CH2NH2 middot HCl FW 11361

HSNH2

bull HCl

ge98 (titration)M6500-25G 25 g

M6500-100G 100 g

34-Dichloro iso cou marin

34-DCI 34-Dichloro-2-benzo pyran-1-one [51050-59-0] C9H4Cl2O2 FW 21503

O

ClCl

OEffective concentration 5-100 μM

Half-life = 20 min at pH 75

Serine protease inhibitor Active towards a wide range of serine proteases including granzymes Not active toward β-lactamases

D7910-5MG 5 mg

D7910-10MG 10 mg

D7910-25MG 25 mg

Dichloro methyl ene diphos pho nic acid disodium salt

Clodronic acid disodium salt Cl2MDP DMDP [22560-50-5] CH2O6Cl2Na2P2 FW 28886 P ONa

O

OHP

Cl

ClHO

ONaO

Analog of pyrophosphate ion that inhibits the osteoclastic activity leading to bone resorption and osteoporosis The compound is used in cancer research especially in skeletal metastases and breast carcinoma12 When entrapped in liposomes it is used for macrophage-selective depletion (macrophage ldquosuiciderdquo technique) especially in spleen and liver3 Found also to inhibit collagenase and matrix metalloproteinase14

Lit cited 1 Lipton A Cancer 80 1668 (1997) 2 Fleisch HA Ann Med 29 55 (1997) 3 Van Rooijen N J Immunol Methods 124 1 (1989) 4 Teronen O et al Calcif Tissue Int 61 59 (1997)

D4434-1G 1 g

Diethyl enetri amine penta acetic acid

DTPA NN-Bis(2-[bis(carboxy methyl)amino]ethyl)gly cine DETAPAC Com plex onereg V Pentetic acid Penta(carboxy methyl) diethyl enetri amine (Carboxy methyl imino)bis (ethylene nitrilo)tetra acetic acid [67-43-6] [(HOOCCH2)2NCH2CH2]2NCH2COOH FW 39335

N

N

O

O

OO N

OH

OH

HO

HO

OH

O

ge99 (titration)Purified

D6518-5G 5 g

D6518-10G 10 g

D6518-50G 50 g

D6518-100G 100 g

Diiso pro pyl fluoro phos phate

DIFP DFP Diiso pro pyl phos phoro fluor idate Phos phor ic acid diiso pro pyl ester fluoride [55-91-4] [(CH3)2CHO]2POF FW 18415

CH3 CHCH3

O PO

FO CH

CH3

CH3

Potent inhibitor of serine proteases such as trypsin and chymotrypsin and of acetylcholinesterase also inhibits cathepsin G cholinesterase coagulation factor Xa leucocyte elastase pancreatic elastase tissue kallikrein plasmin subtilisin and thrombin Inhibition of acetylcholinesterase makes this compound especially toxic Inhibits apoptosis induced by ricin and bacterial toxins

Typically used at a concentration of 010 mM A safer alternative inhibitor for serine protease is phenylmethylsulfonyl fluoride (PMSF)

density 106 gmL 25 degC

D0879-1G 1 g

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

uct Listin

gs

Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

sigma-aldrichcom

Visit sigmacomenzymeexplorer

Visit the Enzyme Explorer Assay Library

Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

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The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

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Page 20: Protease Inhibition and Detection

20 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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E64 AnalogsEnzyme Inhibitor Concentration

Bromelain 10 μM

Calpain 1 amp 2 005 mgml

Cathepsin B (human and rat liver) 10 μM

Cathepsin B1 (squid) 10 μM

Cathepsin H (human liver 10 μM) 10 μM

Cathepsin L (human and rat liver) 10 μM

Cathepsin (rat liver) 28 mM about 82 inhibition

Clostripain 100 μM 81 reversible competitive inhibition

Ficin (10 μM) 10 μM

Papain (10 μM) 10 μM

E-64

l-trans-3-Carboxy oxiran-2-carbo nyl-l-leucylagmatine trans-Epoxy succinyl-l-leucyl amido(4-guani dino)butane N-(trans-Epoxy succinyl)-l-leu cine 4- guani dino butyl amide [66701-25-5] C15H27N5O5 FW 35741

ONH

OOO

HO

CH3

CH3

NH

NHH2N

HN

E-64 is an irreversible potent and highly selective cysteine protease inhibitor E-64 does not react with the functional thiol group of non-protease enzymes such as l-lactate dehydrogenase or creatine kinase E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors leupeptin and antipain The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases such as papain actinidase and cathepsins B H and L to form a thioether linkage E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition it is permeable in cells and tissues and has low toxicity

E-64 is soluble in water A 20 mgml solution can be prepared in water (heat may be needed) A suggested stock solution is a 1 mM aqueous solution The effective concentration for use as a protease inhibitor is 1 to 10 μM Aqueous stock solutions are stable for months at -20 degC Diluted solutions are stable for days at neutral pH E-64 is stable from pH 2-10 but is unstable in ammonia or in HCl E-64 is also soluble in DMSO a 10 mM solution can be prepared in dry DMSO and stored at -20 degC Subsequent dilutions were in culture medium Solutions for injection were prepared by dissolving E-64 in 09 sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 09 sodium chloride (after adjusting the pH to 70 with acetic acid)

E3132-1MG 1 mg

E3132-5MG 5 mg

E3132-10MG 10 mg

E3132-25MG 25 mg

E3132-50MG 50 mg

E-64c(2S3S)-trans-Epoxy succinyl-l-leucyl amido-3-methyl butane [76684-89-4] C15H26N2O5 FW 31438Cysteine protease inhibitor membrane-impermeable calpain inhibitor

E0514-1MG 1 mg

E0514-5MG 5 mg

E-64d

EST (2S3S)-trans-Epoxy-succinyl-l-leucyl amido-3-methyl butane ethyl ester [88321-09-9] C17H30N2O5 FW 34243

C C

O

C NH CH

CH2

CH

CH3 CH3

C

O

NH CH2CH2CHCH3

O

H

C HO

O

CH3CH2

CH3

Cysteine protease inhibitor Synthetic analog of E-64 membrane-permeable calpain inhibitor

E8640-250UG 250 μg

E8640-1MG 1 mg

Elastatinal

(2S)-2-[(4S)-2-Amino-1456-tetra-hydro4-pyrimi dinyl]-N-[[[(1S)-1-carboxy-3-methyl butyl]amino] carbo nyl]glycyl-N1-[(1S)-1-methyl- 2-oxo ethyl-l-glutam amide [51798-45-9] C21H36N8O7 FW 51256

H3CHN

HN

NH

HN

HCH3 O

O

O

O

NH

NH

NH

CH3

OHO

O NH2

H

Potent irreversible elastase inhibitor

~50E0881-5MG 5 mg

E0881-10MG 10 mg

E0881-25MG 25 mg

Epiamastatin hydrochloride

[(2R3R)-3-Amino-2-hydroxy-5-methyl-hexa noyl]-Val-Val-Asp hydrochloride [100992-59-4] C21H38N4O8 FW 47455

NH

HN

O

OOH

NH2

bull HCl

H3C

CH3

NH

OHO

O

O

OH

H3C CH3

CH3H3C

Metallo-protease inhibitor with selectivity for aminopeptidases

ge97 (HPLC)E3389-1MG 1 mg

E3389-5MG 5 mg

q Ethylenediaminetetraacetic acid

Edathamil

Ethylene di amine tetra acetic acid disodium salt dihydrate

Sequestrene Na2 Disodium ethylene di amine tetra acetate dihydrate Edetate disodium salt dihydrate EDTA disodium salt [6381-92-6] C10H14N2Na2O8 middot 2H2O FW 37224

ORN

NRO

O

O

RO

O

OR

OR = 2 H and 2 Na

bull H2O

bull H2O

Chelator of divalent cations Inhibits enzymes such as metalloproteases that require divalent cations for activity

reagent grade ~99 (titration)Zinc-dependent metalloproteinases as well as other proteases that are stabilized by calcium can be effectively inhibited by chelation of divalent metal ions with EDTA Other chelators such as EGTA specific for calcium and 110-phenanthroline highly specific for zinc can be used to target these two different types of proteases

ED2SS-50G 50 g

ED2SS-100G 100 g

ED2SS-250G 250 g

ED2SS-500G 500 g

ED2SS-1KG 1 kg

ED2SS-25KG 25 kg

ED2SS-5KG 5 kg

Ethylene di amine tetra acetic acid tripotassium salt dihydrate

Tri potas sium ethylene di amine tetra acetate dihydrate EDTA tripotassium salt [65501-24-8] (KOOCCH2)2NCH2CH2N(CH2COOK)CH2COOH middot 2H2O FW 44254

NN

KO

O

OK

OO

HO

OKO

bull 2H2O

Used to eliminate inhibition of enzyme catalyzed reactions due to traces of heavy metals

98 (titration)E0270-25G 25 g

E0270-100G 100 g

Ethylenediaminetetraacetic acid p

Our Innovation Your Research mdash Shaping the Future of Life Science 21

Prod

uct Listin

gs

Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

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Features over 600 detailed procedures for measuring enzyme activity and related metabolites

The Library is the result of over ten years of in-house work on analytical procedures developed

by Sigma-Aldrichreg scientists

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

gs

Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

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Characterization

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amp shRNA

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

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Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 21: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 21

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Ethylene glycol-bis(2-amino ethyl ether)-NNNprimeNprime-tetra acetic acid

Egtazic acid EGTA Glycol ether di amine tetra acetic acid Ethylene-bis(oxy ethylene nitrilo)tetra acetic acid Cheltrade-DE [67-42-5] [-CH2OCH2CH2N(CH2CO2H)2]2 FW 38035

NO

ON

O

OH

HO

O

O

HO

OH

O

A chelating agent useful for the determination of calcium in the presence of magnesium

ge970The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the ELITE level

E4378-10G 10 g

E4378-25G 25 g

E4378-100G 100 g

E4378-250G 250 g

E4378-500G 500 g

E4378-1KG 1 kg

N-Ethyl male imide

NEM [128-53-0] C6H7NO2 FW 12513 N

CH3

O O

Augments currents from native M-channels in sympathetic neurons and acts as an opener for KCNQ2 KCNQ4 and KCNQ5 channels

Sulfhydryl alkylating agent that inactivates NADP-dependent isocitrate dehydrogenase and many endonucleases

Reagent for the covalent modification of cysteine residues in proteins

crystalline ge98 (HPLC)NEM is an inhibitor of cysteine proteases such as calpain NEM binds stoichiometrically and is also used as a reagent for sulfhydryl deter minations with a sensitivity to 01 mM

The adsorption spectrum has a maximum at approximately 300 nm which disappears as NEM reacts with sulfhydryl groups

E3876-5G 5 g

E3876-25G 25 g

E3876-100G 100 g

NEM gives a clear solution in ethanol at 50 mgml NEM dissolves in water (gt50 mg in 4 ml) however aqueous solutions are unstable The rate of hydrolysis is pseudo-first order and significantly dependent on pH The half-lives of solutions at different pH were reported Preparation of fresh solutions for each usage is recommended The solution concentration should be essentially constant for approximately three hours at room temperature at or below pH 70

pH Half-life (hrs)

65 160

7 48

75 9

8 17

825 07

85 033

Foroxymithine microbial

[100157-28-6] C22H37N7O11 FW 57557

ge85 (HPLC)F5017-1MG 1 mg

Gabexate mesylate

4-[[6-[(Amino imino methyl)amino]-1-oxo hexyl]oxyl]benzoic acid ethyl ester mesylate salt FOY Megacart [56974-61-9] C16H23N3O4 middot CH4O3S FW 41748

SO

OOHH3C

O

ONH

NH2

NH

O

O

H3C

solidSerine protease inhibitor

G2417-5MG 5 mg

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Features over 600 detailed procedures for measuring enzyme activity and related metabolites

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

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Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 22: Protease Inhibition and Detection

22 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Glu-Val-Asn-[(2R4S5S)-5-amino-4-hydroxy-27-dimethyl octa noyl]-Ala-Glu-PheOM99-2 [314266-76-7] C41H64N8O14 FW 89299Inhibitor of brain β-secretase

gt93 (HPLC) solidG8291-5MG 05 mg

Gly-Gly-Tyr-Arg[70195-20-9] C19H29N7O6 FW 45148

ge97 (HPLC)Useful for affinity chromatography

G5386-100MG 100 mg

Hirudin from leeches[8001-27-2]The anticoagulant hirudin is the most potent natural inhibitor of both soluble and clot-bound thrombin It binds thrombin with high affinity covering more than 20 of its surface area and occluding both the active site and exosite I (fibrinogen and PAR recognition site) Hirudin blocks thrombus growth and platelet activation It is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration

Hirudin is a 7 kDa acidic protein containing 65 amino acid residues that has an isoelectric point of 35-40 It is not glycosylated and lacks tryptophan arginine and methionine residues It is chemically stable between pH 2-12 and at temperatures up to 80degC

One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

lyophilized powder activity ge1500 ATUmg protein (ATU = antithrombin units)

H7016-10UN 10 units

H7016-100UN 100 units

H7016-500UN 500 units

H7016-2KU 2000 units

lyophilized powder activity 300-1500 ATUmg protein (ATU = antithrombin units)

Lyophilized from 0045 M NaCl and 002 M Tris buffer pH 75

H7380-100UN 100 units

H7380-500UN 500 units

H7380-2KU 2000 units

[Lys47]Hirudin (HV2) leech[120300-65-4]

recombinant expressed in Saccharomyces cerevisiae activity ge7000 ATUmg protein (ATU = antithrombin units)One unit (ATU) will neutralize one NIH unit of thrombin at 37 degC based on direct comparison to an NIH thrombin reference standard

Protein determined by Lowry

H0393-100UN 100 units

H0393-500UN 500 units

Histatin 5Asp-Ser-His-Ala-Lys-Arg-His-His-Gly-Tyr-Lys-Arg-Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr [104339-66-4] C133H195N51O33 FW 303629

ge97 (HPLC)A human salivary peptide that inhibits the protease of Bacteroides gingivalis as well as clostripain

H6027-1MG 01 mg

Ile-Pro-IleDiprotin A [90614-48-5] C17H31N3O4 FW 34145Inhibitor of dipeptidyl peptidase IV Inhibits entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines

ge97 (HPLC)I9759-5MG 5 mg

I9759-25MG 25 mg

Isoamyl phos pho nyl-Gly-Pro-Ala dipotassium salt

Reported inhibitor of clostridial collagenase

gt95 (NMR TLC) lyophilized powderI7029-5MG 5 mg

Lactacystin

[133343-34-7] C15H24N2O7S FW 37643

NH

O

H3C HO

S OH

OH

H3C CH3

O

HN

O

O

CH3

ge90 (TLC)Cell-permeable and irreversible proteasome inhibitor (Ki = 4 nM) Inhibits NF-κB activation (IC50 = 10 μM) Induces neurite outgrowth in neuro 2A mouse neuroblastoma cells

L6785-2MG 02 mg

L-Leu cine thiol oxidized dihydrochlorideDithio bis(2-amino-4-methyl pentane) [88264-65-7] C12H28N2S2 middot 2HCl FW 33742

L8397-1MG 1 mg

L8397-5MG 5 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

gs

Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

Donrsquot Have Time

Sigmalsquos commitment to life science research extends far beyond quality products

Find out how we can help at sigmacomlifescienceservices with our array of services that will accelerate your research with a new level of confidence and convenience

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 23: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 23

Prod

uct Listin

gs

Leupeptin

N

O

O

N

O

N

O

H

H

H

H

CH2CH(CH3)2

CH2CH(CH3)2

CH2CH2CH2

CH3

CNH

NH

NH2

Leupeptin Protease Inhibitor Specificity

Enzyme Ki

Acrosin Ki = 210 nM

Calpains Ki = 10-430 nM

Cathepsin B Ki = 41 nM

Chymotrypsin Ki = 11 mM

Histolysin Ki = 43 nM

Plasmin Ki = 34 mM

Trypsin Ki = 35 nM

q Leupeptin

Inhibitor of serine and cysteine proteases Inhibits plasmin trypsin papain and cathepsin B No inhibition found with pepsin cathepsins A and D thrombin or α-chymotrypsin Effective concentration 10-100 μM

Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution Purity determined using three main peaks Majority of contaminating peptide is racemized leupeptin

Leupeptin hydrochlorideAcetyl-Leu-Leu-Arg-al hydrochloride [24125-16-4] C20H38N6O4 middot HCl FW 46301

ge90 (HPLC)L9783-5MG 5 mg

L9783-25MG 25 mg

L9783-100MG 100 mg

ge70 (HPLC)L0649-5MG 5 mg

L0649-25MG 25 mg

L0649-100MG 100 mg

Leupeptin hemisulfate salt

N-Acetyl-l-leucyl-l-leucyl-l-argininal hemisulfate salt Acetyl-Leu-Leu-Arg-al [103476-89-7] C20H38N6O4 middot 12H2SO4 FW 47559

ge90 (HPLC)The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

L2884-5MG 05 mg

L2884-1MG 1 mg

L2884-5MG 5 mg

L2884-10MG 10 mg

L2884-25MG 25 mg

L2884-50MG 50 mg

L2884-100MG 100 mg

L2884-250MG 250 mg

ge85 (HPLC)L8511-5MG 5 mg

L8511-10MG 10 mg

L8511-25MG 25 mg

L8511-50MG 50 mg

L8511-100MG 100 mg

L8511-250MG 250 mg

BioUltra ge98 (HPLC) 8

L5793-5MG 5 mg

L5793-25MG 25 mg

L5793-100MG 100 mg

Leupeptin trifluoroacetate saltAcetyl-Leu-Leu-Arg-al trifluoroacetate salt [147385-61-3] C20H38N6O4 middot xC2HF3O2 FW 42655 (FB)

ge90 (HPLC)L2023-1MG 1 mg

L2023-5MG 5 mg

L2023-10MG 10 mg

L2023-25MG 25 mg

L2023-50MG 50 mg

L2023-100MG 100 mg

Leupeptin p

Cloning amp

Transfection

Protein Expression

Services

Mass Spec ampProtein

Characterization

Biomarker Support Services

Custom siRNA

amp shRNA

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Sigmalsquos commitment to life science research extends far beyond quality products

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24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

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isti

ng

s

Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 24: Protease Inhibition and Detection

24 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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α2-Macro globulin from human plasma

α2-M

α2-Macroglobulin is found abundantly in plasma and interstitial fluids The protease-α2-M balance plays an important role in mediating inflammation-associated tissue destruction Serum levels of α2-M and protease-α2-M complexes are increased in patients with sepsis emphysema periodontitis rheumatoid arthritis and other inflammatory diseases It is hypothesized that the oxidant inactivation of α2-M contributes to tissue destruction during inflammation α2-M has been implicated as a genetic risk factor for late-onset Alzheimerrsquos disease Activated α2-M enhances the clearance of soluble αβ-amyloid via low-density lipoprotein receptor-related protein in cortical neurons but has no effect on secreted or full-length APP levels

Inhibits all classes of endoproteases by forming a 11 complex with the protease When the protease cleaves the macroglobulin ldquobaitrdquo sequence the macroglobulin rearranges and traps the protease

Plasma from each donor has been tested and found negative for antibody to HIV-1HIV-2 antibody to HCV and HbSAg

Bait Region

Inhibitory Regions

Signal Peptide (removed)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

= N-linked Glycans

α2-Macroglobulin(Human Sequence)

= Disulfides = Interchain Disulfides

Isoglutamyl-lysine isopeptides

α2-Macroglobulin is a tetramer with four identical subunits with molecular weights of 179 kDa Upon binding to a protease the 179 kDa subunit is cleaved into two 85 kDa fragments

BioUltra lyophilized powder ge98 (SDS-PAGE)Lyophilized from 002 M Tris 013 M glycine pH 80 and 008 M trehalose

Package size based on protein content

One mg of α2-macroglobulin will inhibit a minimum of 10 μg trypsin with an activity of 10000 BAEE unmg protein

65142

20

2429

36

45

5566

97116

200

Co

mp

etitor

Sigm

a

SDS-PAGE (7) of α2-macroglobulin (Cat No M6159) under reducing conditions and stained with Coomassie blue

M6159-1MG 1 mg

M6159-5MG 5 mg

M6159-10MG 10 mg

N-(Methoxy succinyl)-Ala-Ala-Pro-Val-chloro methyl keto ne

N-(Methoxy succinyl)-l-alanyl-l-alanyl-l-prolyl-l-valine chloro methyl keto ne [65144-34-5] C22H35ClN4O7 FW 50299

Ala Ala Pro Val Cl

OH3CO

O

Inhibitor of human granulocyte elastase

M0398-5MG 5 mg

M0398-10MG 10 mg

M0398-25MG 25 mg

N-Methoxy succinyl-Val-Homo-Phe-fluoro methyl keto neMu-Val-HPhe-FMK C21H30N3O4F FW 40748Calpain inhibitor peptide

ge95 (TLC) powderM5439-3MG 3 mg

Pepstatin A

[26305-03-3] C34H63N5O9 FW 68589

CH3

OH

HN

NH

CH3

CH3

OOH

O

CH3OH

CH3

OHN

NH

CH3

H3CO

CH3

O

HN

OCH3H3C

H3C

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases) It forms a 11 complex with proteases such as pepsin renin cathepsin D bovine chymosin and protease B (Aspergillus niger) The inhibitor is highly selective and does not inhibit thiol proteases neutral proteases or serine proteases Solubilized γ-secretase and retroviral protease are also inhibited by Pepstatin A It has been used to characterize proteases from several sources

Pepstatin can be dissolved at 1 mgml in 10 (vv) acetic acid in methanol (91 methanolacetic acid) The inclusion of acetic acid may be necessary to dissolve this peptide in methanol or DMSO It has been dissolved at 1 to 2 mgml in ethanol but heat may be required for complete dissolution Solutions of Pepstatin A can be heated as high as 60 degC without any decomposition of the peptide

Stock solutions at 1 mgml are stable at least a week at 4 degC A 1 mM solution in methanol or DMSO is stable for months at -20 degC If solutions become darker yellow the reagent is hydrolyzing

A working concentration of 1 μM is stable for at least one day at room temperature

ge90 (HPLC)P5318-5MG 5 mg

P5318-25MG 25 mg

ge75 (HPLC)P4265-1MG 1 mg

P4265-5MG 5 mg

P4265-25MG 25 mg

P4265-100MG 100 mg

P4265-250MG 250 mg

Phenyl meth ane sul fonyl fluoride

PMSF α-Tolu ene sul fonyl fluoride Phenyl methyl sul fonyl fluoride Benzyl sul fonyl fluoride [329-98-6] C7H7FO2S FW 17419

S FO

O

ge985 (GC)Inhibits serine proteases such as trypsin and chymotrypsin Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Not as effective or as toxic as DFP Effective concentration 01-1 mM Half-life = 1 hr at pH 75

P7626-250MG 250 mg

P7626-1G 1 g

P7626-5G 5 g

P7626-25G 25 g

P7626-100G 100 g

α2-Macroglobulin (Human Sequence)

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

uct Listin

gs

Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

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uct Listin

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Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 25: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 25

Prod

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Phos phor amido n disodium salt

N-(α-Rhamno pyran osyl phos phono)-l-leucyl-l-trypto phan disodium salt N-(α-Rhamno pyran osyl oxy hydroxy phos phinyl)-Leu-Trp disodium salt [119942-99-3] C23H32N3Na2O10P FW 58747

POO

HN

HN

HNO-

O-O

O

OOH

OHHO

Na+

Na+H3C

H3C

CH3

ge97 (TLC)Solubility testing at 10 mgml in water yields a clear colorless to light yellow-green solution The product is also soluble in methanol and DMSO less soluble in ethanol and ethyl acetate insoluble in benzene hexane and chloroform

Solutions can be stored in aliquots at -20 degC with an expected shelf life of at least one month

Potent inhibitor of thermolysin and other bacterial metallo-endopeptidases Strongly inhibits mammalian enkephalinase Inhibits some metallo-endopeptidases Does not inhibit trypsin papain chymotrypsin or pepsin Weakly inhibits collagenase Effective concentration 1-10 μM

R7385-5MG 05 mg

R7385-1MG 1 mg

R7385-5MG 5 mg

R7385-10MG 10 mg

R7385-25MG 25 mg

Procathepsin B Fragment 26-50 ratAla-Gly-Arg-Asn-Phe-Tyr-Asn-Val-Asp-Ile-Ser-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C123H198N34O33S FW 271316Cathepsin B inhibitor

ge95 (HPLC)P0350-5MG 05 mg

Procathepsin B Fragment 36-50 ratSer-Tyr-Leu-Lys-Lys-Leu-Cys-Gly-Thr-Val-Leu-Gly-Gly-Pro-Lys-NH2 C71H123N19O18S FW 156292Cathepsin B inhibitor

ge95 (HPLC)P0475-5MG 05 mg

gt Protease Inhibitor Cocktails and Tablets see Protease Inhibitor Cocktails and Tablets Page 6

β-Secretase inhibitorKTEEISEVN[Statine]VAEF C73H118N16O27 FW 165181

ge97 (HPLC) powderS4562-1MG 1 mg

γ-Secretase inhibitorC33H57F2N5O9 FW 70583

ge90 (TLC) powderA reversible γ-secretase inhibitor

S2188-1MG 1 mg

N-Succinyl-L-proline[63250-32-8] C9H13NO5 FW 21520

S6633-100MG 100 mg

S6633-1G 1 g

Nα-Tosyl-L-lysine chloro methyl keto ne hydrochloride

(3S)-1-Chloro-3-tosyl amido-7-amino-2- hepta none hydrochloride Tosyl-l-lysyl- chloro meth ane hydrochloride (3S)-7-Amino- 1-chloro-3-tosyl amino-2-hepta none hydrochloride TLCK[4272-74-6] C14H21ClN2O3S middot HCl FW 36931

O

NH

H

SO

O

H3C

HCl

NH2

Cl

ge96 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis in cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T7254-100MG 100 mg

T7254-500MG 500 mg

T7254-1G 1 g

T7254-5G 5 g

N-p-Tosyl-L-phenyl ala nine chloro methyl keto ne

TPCK (S)-1-Chloro-3-tosyl amido-4-phenyl-2-butanone Tosyl-l-phenyl alanyl-chloro meth ane (S)-1-Chloro-4-phenyl-3-tosyl amido-2-butanone [402-71-1] C17H18ClNO3S FW 35185

CHNH

CO

CH2Cl

S

CH3

OO

CH2

ge97 (TLC) powderBlocks the LPS- or cytokine-induced activation of nuclear factor κB (NFκB) which in turn blocks the induction of iNOS and COX-2 transcription Blocks activation of pp70s6k by all mitogens Blocks apoptosis cell lines by inhibiting the processing of caspases in some cell lines and to some stimuli

T4376-100MG 100 mg

T4376-250MG 250 mg

T4376-1G 1 g

T4376-5G 5 g

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

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0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 26: Protease Inhibition and Detection

26 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Trypsin InhibitorsNatural trypsin Inhibitors also known as serine protease inhibitors (serpins) are the largest and most diverse family of protease inhibitors Serpins control the activation and catabolism of proteins by the inhibition of serine proteases in vivo

There are four primary natural sources of trypsin inhibitors bovine pancreas ovomucoid soybean and lima bean Each inhibitor acts as a competitive substrate analog and binds with its serine protease to form an inactive complex therefore rendering the protease inactive

This process allows the serpin (trypsin inhibitor) to stop the proteolytic activity of the serine protease when its function is no longer necessary

Trypsin inhibitors provide unique processes depending on their source For example inhibitors in the seeds of legumes (soybean and lima bean) act as a feeding deterrent for insects by disrupting midgut proteases This natural function is being expanded upon in the development of insect resistant transgenic plants Soybean inhibitors have also been found to contribute to pancreatic hypertrophy in rats again providing a feeding deterrent The Bowman-Birk soybean inhibitor is being studied as a cancer preventive agent

Trypsin-chymotrypsin inhibitor from Glycine max (soybean)Bowman-Birk Inhibitor [37330-34-0]

10 20 30 40 50 60 70 80 90 100 110

Trypsin binding site

Chymotrypsin binding site

7 Disulfides

Signal Peptide

Propeptide

Trypsin Inhibitor (Bowman-Birk)(soybean)

Trypsin Inhibitor (Bowman-Birk) (soybean)

lyophilized powderBowman Birk protease inhibitor prevents radiation-induced carcinogenesis by a reduction of incorrect DNA repairs resulting in a reduced amount of dicentric chromosomes

Lyophilized powder containing phosphate buffer salts pH 76

One trypsin unit = ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

1 mg protein will inhibit ge05 mg trypsin with activity of ~10000 BAEE units per mg protein or ge10 mg chymotrypsin with activity of ~40 BTEE units per mg protein

T9777-10MG 10 mg

T9777-25MG 25 mg

T9777-50MG 50 mg

T9777-100MG 100 mg

T9777-500MG 500 mg

q Trypsin inhibitor

[9035-81-8]

Trypsin inhibitor from bovine pancreasBPTI

10 20 30 40 50 60 30 0 70 80 90 100 10 20 30 40 50 60 70 80 90 100

Trypsin binding site

3 Disulfides

Signal Peptide

Trypsin Inhibitor (bovine pancreas)

Propeptide Propeptide

Kunitz domain

Trypsin Inhibitor (bovine pancreas)

Type I-P essentially salt-free lyophilized powderDerived from New Zealand-sourced pancreas

Prepared by method of Kunitz and Northrup J Gen Physiol 19 991 (1936)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg of trypsin inhibitor will inhibit greater than 15 mg of trypsin with activity of approximately 10000 BAEE units per mg protein

Protein determined by biuret

T0256-1MG 1 mg

T0256-5MG 5 mg

T0256-10MG 10 mg

T0256-25MG 25 mg

Trypsin inhibitor from chicken egg white

Ovo muco id

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

9 Disulfides

Signal Peptide

Trypsin Inhibitor (chicken egg white)

N-Linked Glycans

Kazal-like domains

Trypsin Inhibitor (chicken egg white)

Type II-O Partially purified ovomucoid containing ovoinhibitorOne mg (biuret) will inhibit 08-12 mg of tryspin with activity of approx 10000 BAEE units per mg protein May inhibit le03 mg of chymotrypsin with activity of approx 40 BTEE units per mg protein

T9253-250MG 250 mg

T9253-500MG 500 mg

T9253-1G 1 g

T9253-5G 5 g

Type III-O (free of ovoinhibitor)One milligram of product will inhibit 10 - 20 mg of trypsin with activity of ~10000 BAEE unitsmg protein

T2011-250MG 250 mg

T2011-500MG 500 mg

T2011-1G 1 g

T2011-5G 5 g

Trypsin Inhibitor (Bowman-Birk) (soybean)

Trypsin Inhibitor (bovine pancreas)

Trypsin Inhibitor (chicken egg white)

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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s

Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

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Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

sigma-aldrichcom

LFL71112-508361

0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 27: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 27

Prod

uct Listin

gs

Trypsin inhibitor from Glycine max (soybean)

SBTI

Soybean trypsin inhibitor inhibits trypsin and to a lesser extent chymotrypsin and plasmin It forms a 11 stoichiometric complex with trypsin Upon formation of this complex trypsin may cleave a single arginine-isoleucine bond in the inhibitor Dissociation of this complex may yield the modified form or the native inhibitor At the optimal pH for trypsin binding (pH 80) the association constant is ge 10x108

Trypsin inhibitor is soluble in water and phosphate buffers at concentrations of 10 mgml or higher Solutions at higher concentrations may be hazy and have a yellow to amber color

20 40 60 80 100 120 140 160 180 200

Trypsin binding site

2 Disulfides

Signal Peptide

Trypsin Inhibitor (Kunitz)(soybean)

Propeptide

Trypsin Inhibitor (Kunitz) (soybean)

Type I-S lyophilized powderChromatographically prepared

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One chymotrypsin unit will hydrolyze 10 μmole of BTEE per min at pH 78 at 25 degC

One mg will inhibit 10-30 mg of trypsin with activity of approx 10000 BAEE units per mg protein

Protein determined by biuret

The Enhanced Quality Profile (EQP) is an SAFC program characterizing a productrsquos quality traits in a 5-level system to meet the requirements of our manufacturing customers This product is in the PREMIUM level

T9003-25MG 25 mg

T9003-100MG 100 mg

T9003-250MG 250 mg

T9003-500MG 500 mg

T9003-1G 1 g

T9003-5G 5 g

powder cell culture testedAfter trypsinizing cells resuspend cells in 1 mL trypsin inhibitor solution (1mgml) for every mL of trypsin solution used for dissociation Centrifuge the cell suspension at 1000 rpm for 5 minutes A cell pellet should form Remove as much of the trypsin inhibitor solution as possible and resuspend the pellet in serum-free medium Culture cells as desired

Type I-S

activity (One mg will inhibit 1-3 mg of trypin with activity of approx 10000 BAEE units per mg protein)

Chromatographically prepared

One trypsin unit = ΔA253 of 0001 per minute with Nminusα-benzoyl-l-arginine ethyl ester (BAEE) as substrate at pH 76 at 25 degC Reaction volume = 32 mL (1 cm light path)

Protein determined by biuret

T6522-25MG 25 mg

T6522-100MG 100 mg

T6522-5X100MG 5 times 100 mg

T6522-250MG 250 mg

T6522-500MG 500 mg

T6522-1G 1 g

T6522-5G 5 g

lyophilized powderType II-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit minimum 10 mg of trypsin with activity of approx10000 BAEE units per mg protein

T9128-500MG 500 mg

T9128-1G 1 g

T9128-5G 5 g

T9128-10G 10 g

1 solution in water (contains no buffers)Prepared From Type I-S

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

Protein determined by biuret

T9008-5ML 5 mL

1 times solution cell culture testedContains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum

Optimized for passage of endothelial cell cultures

Prepared in Dulbeccorsquos Phosphate Buffered Saline without calcium and magnesium

Endotoxin tested

T6414-100ML 100 mL

T6414-6X100ML 6 times 100 mL

Trypsin inhibitor from Phaseolus limensis (lima bean)

Trypsin Inhibitor from lima beans

Monomer has an apparent molecular weight of approx 9000 but undergoes a concentration and pH-dependent dimerization

Type II-L crude powderPrepared by a modification of the method of Tauber H et al J Biol Chem 179 1155 (1949) (modified)

One trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 mL 1 cm light path

One mg (biuret) will inhibit a minimum of 08 mg of trypsin with activity of approx 10000 BAEE units per mg protein

T9378-100MG 100 mg

T9378-500MG 500 mg

T9378-1G 1 g

Trypsin inhibitor from turkey egg white

Type II-TOne trypsin unit will produce a ΔA253 of 0001 per min with BAEE as substrate at pH 76 at 25 degC reaction volume 32 ml 1 cm light path

One mg (biuret) will inhibit 09-13 mg of trypsin with activity of approx 10000 BAEE units per mg protein or 04-10 mg of α-chymotrypsin with activity of approx 40 BTEE units per mg protein

T4385-250MG 250 mg

T4385-1G 1 g

T4385-5G 5 g

Trypsin inhibitor p

Trypsin Inhibitor (Kunitz) (soybean)

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

du

ct L

isti

ng

s

Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e D

etec

tio

n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

sigma-aldrichcom

LFL71112-508361

0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

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Page 28: Protease Inhibition and Detection

28 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

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ct L

isti

ng

s

Urinary Trypsin Inhibitor FragmentArg-Gly-Pro-Cys-Arg-Ala-Phe-Ile Bikunin[164859-77-2] C40H66N14O9S FW 91911

ge95 (HPLC)Blocks the metastasis of human ovarian cell line (HRA) without affecting their proliferation

Fragment of domain II of the urinary trypsin inhibitor (UTI) corresponds to active sequence of UTI for plasmin inhibition

U4751-1MG 1 mg

Z-Ala-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-AEVD-FMK C28H39N4O10F FW 61063Caspase inhibitor

solidge90 (TLC)

C8484-1MG 1 mg

Z-Asp(O-Me)-Glu(O-Me)-Val-Asp(O-Me) fluoro methyl keto neZ-DEVD-FMK C30H41N4O12F FW 66866A cell-permeable inhibitor of caspase 3 6 7 8 10 which exhibits competitive and irreversible inhibition

ge90 (TLC)C0605-3MG 3 mg

Z-Asp(OMe)-Gln-Met-Asp(OMe) fluoro methyl keto neZ-DQMD-FMK dimethyl ester C29H40N5O11FS FW 68572

powderA cell-permeable inhibitor of caspase 3 and 6 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C0480-1MG 1 mg

Z-Gly-Leu-Phe-chloro methyl keto neZ-GLF-CMK C26H32N3O5Cl FW 50200Cathepsin G inhibitor

ge90 (TLC) solidC9984-10MG 10 mg

Z-Ile-Glu(O-ME)-Thr-Asp(O-Me) fluoro methyl keto neC30H43N4O11F FW 65468

Z-IETD-FMK ge90 (TLC) powder

Cell-permeable inhibitor of caspase-8 which exhibits competitive and irreversible inhibition

C1230-1MG 1 mg

Z-Leu-Glu(O-ME)-His-Asp(O-Me) fluoro methyl keto neC32H42FN6O10 FW 68971

Z-LEHD-FMK powder

A cell-permeable inhibitor of caspase-9 which exhibits competitive and irreversible inhibition

ge90 (TLC)

C1355-1MG 1 mg

Z-Leu-Glu(OMe)-Glu(OMe)-Asp(OMe)-fluoro methyl keto neZ-LEED-FMK C32H45FN4O12 FW 69672

solidCell permeable Caspase-13 inhibitor

ge90 (TLC)

C8859-1MG 1 mg

Z-Leu-Glu(OMe)-Thr-Asp(OMe)-fluoro methyl keto neZ-LETD-FMK C30H44N4O11F FW 65569

solidCell permeable caspase-8 inhibitor

ge90 (TLC)

C8734-1MG 1 mg

Z-Leu-Glu(OMe)-Val-Asp(OMe)-fluoro methyl keto neZ-LEVD-FMK Caspase 1 inhibitor C31H45N4O10F FW 65271Cell permeable caspase-1 inhibitor

ge90 (TLC) solidC9484-3MG 3 mg

Z-Leu-Leu-Leu-fluoro methyl keto neZ-LLL-FMKC27H42N3O5F FW 50764

ge90 (TLC) solidCalpain Inhibitor

C8984-3MG 3 mg

Z-Leu-Leu-Phe-CHOZ-LLF-CHO Cbz-Leu-Leu-Phe-CHO C29H39N3O5 FW 50964Cell permeable proteasome inhibitor acting on the chymotrypsin-like activity of the multicatalytic proteinase complex

ge95 (HPLC) solidManufactured for Sigma by Boston Biochem Inc

C0358-1MG 1 mg

Z-Phe-Phe-fluoro methyl keto neZ-FF-FMKC27H27N2O4F FW 46251

ge98 (TLC) solidCathepsin B and Cathepsin L inhibitor

C9109-3MG 3 mg

Z-Val-Asp(O-Me)-Val-Ala-Asp(O-Me) fluoro methyl keto neZ-VDVAD-FMK C32H46N5O11F FW 69573

powder ge90 (TLC)A cell-permeable inhibitor of caspase-2 which exhibits competitive and irreversible inhibition

C1605-1MG 1 mg

Z-Val-Glu(O-Me)-Ile-Asp(O-Me) fluoro methyl keto neZ-VEID-FMK C31H45N4O10F FW 65271

powderA cell-permeable inhibitor of caspase-6 which exhibits competitive and irreversible inhibition

C1730-1MG 1 mg

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e D

etec

tio

n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

sigma-aldrichcom

LFL71112-508361

0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom

OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052

Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice

DevelopmentBulk Manufacturing Inquiries (800) 244-1173

Sigma-Aldrich3050 Spruce StreetSt Louis MO 63103 USAsigma-aldrichcom

Accelerating Customersrsquo

Success through Innovation and

Leadership in Life Science

High Technology and Service

ArgentinaSIGMA-ALDRICH DE ARGENTINA SA Free Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD Free Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

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BelgiumSIGMA-ALDRICH NVSAFree Tel 0800 14747 Free Fax 0800 14745 Tel (+32) 3 899 13 01 Fax (+32) 3 899 13 11

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Czech RepublicSIGMA-ALDRICH spol s r o Tel (+420) 246 003 200 Fax (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK AS Tel (+45) 43 56 59 10 Fax (+45) 43 56 59 05

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FranceSIGMA-ALDRICH CHIMIE Sagraverl Free Tel 0800 211 408Free Fax 0800 031 052Tel (+33) 474 82 28 00 Fax (+33) 474 95 68 08

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HungarySIGMA-ALDRICH Kft Ingyenes telefonszaacutem 06 80 355 355 Ingyenes fax szaacutem 06 80 344 344 Tel (+36) 1 235 9055 Fax (+36) 1 235 9050

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IrelandSIGMA-ALDRICH IRELAND LTD Free Tel 1800 200 888 Free Fax 1800 600 222Tel +353 (0) 402 20370Fax + 353 (0) 402 20375

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ItalySIGMA-ALDRICH Srl Numero Verde 800 827018 Tel (+39) 02 3341 7310 Fax (+39) 02 3801 0737

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KoreaSIGMA-ALDRICH KOREA Free Tel (+82) 80 023 7111 Free Fax (+82) 80 023 8111 Tel (+82) 31 329 9000 Fax (+82) 31 329 9090

MalaysiaSIGMA-ALDRICH (M) SDN BHDTel (+60) 3 5635 3321 Fax (+60) 3 5635 4116

MexicoSIGMA-ALDRICH QUIacuteMICA SA de CVFree Tel 01 800 007 5300 Free Fax 01 800 712 9920 Tel 52 722 276 1600 Fax 52 722 276 1601

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SingaporeSIGMA-ALDRICH PTE LTDTel (+65) 6779 1200 Fax (+65) 6779 1822

SlovakiaSIGMA-ALDRICH spol s r o Tel (+421) 255 571 562Fax (+421) 255 571 564

South AfricaSIGMA-ALDRICH SOUTH AFRICA (PTY) LTDFree Tel 0800 1100 75 Free Fax 0800 1100 79 Tel (+27) 11 979 1188 Fax (+27) 11 979 1119

SpainSIGMA-ALDRICH QUIacuteMICA SA Free Tel 900 101 376 Free Fax 900 102 028 Tel (+34) 91 661 99 77 Fax (+34) 91 661 96 42

SwedenSIGMA-ALDRICH SWEDEN ABTel (+46) 8 742 4200 Fax (+46) 8 742 4243

SwitzerlandSIGMA-ALDRICH CHEMIE GmbH Free Tel 0800 80 00 80 Free Fax 0800 80 00 81 Tel (+41) 81 755 2828 Fax (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTDFree Tel 0800 717 181 Free Fax 0800 378 785 Tel (+44) 1747 833 000 Fax (+44) 1747 833 313 SAFC (UK) Tel 01202 712305

United StatesSIGMA-ALDRICH PO Box 14508 St Louis Missouri 63178 Toll-Free 800 325 3010 Toll-Free Fax 800 325 5052 Call Collect (+1) 314 771 5750 Tel (+1) 314 771 5765 Fax (+1) 314 771 5757

VietnamSIGMA-ALDRICH PTE LTD VN ROTel (848) 3516 2810 Fax (848) 6258 4238

Internet sigma-aldrichcom

Page 29: Protease Inhibition and Detection

Our Innovation Your Research mdash Shaping the Future of Life Science 29

Protease D

etection

Protease DetectionThe following products are designed for the detection of non-specific proteolytic activity Sigma-Aldrichreg offers a complete line of specific protease substrates such as caspase calpain plasmin and thrombin substrates Visit the Substrate Index in the online Enzyme Explorer at sigma-aldrichcomenzymeexplorer

Fluorescent Protease DetectionNow there is an easy-to-use kit to detect trace amounts of protease or determine total protease activity using the same methodology utilized by Sigmaregrsquos QC department for years Sigma has used this protocol to test thousands of samples for trace contaminating proteolytic activity

The Fluorescent Protease Detection Kit is a complete kit for the detection of primary or trace protease activity Everything you need including control standards buffers and substrate is included For convenience the assay can be performed in either a cuvette or microplate format Each kit contains enough reagents for 200 x 1 mL assays

Protease Fluorescent Detection Kit

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry It is also suitable for detection of trace amounts of protease contamination The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate

Detection limit approx 5 ng trypsin

Supplied with sufficient reagents for up to 200 one mL assays

Tested for detection of all four proteolytic classes (serine apartic cysteine and metalloproteinases)

Add sample to microfuge tube

Add buffer and FITC-casein solution and incubate at 37degC for 60 min

Add TCA solution to stop reaction incubate at 37degC and ppt unreacted substrate

Centrifuge

Remove Supernatant

Transfer dilute and read in a fluorimeter versus standard curve

010000

20000300004000050000600007000080000

0 5 10 15 20 25

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Nanograms of Trypsin

Typical Activity Curve Using Trypsin

0

10000

20000

30000

0 1 2 3 4 5 6 7

Nanograms of Trypsin

Flu

ore

scen

t U

nit

s (4

855

35 n

m)

Sensitivity of the Protease Fluorescent Detection Kit Using Trypsin

Protease Protease ClassDetection Limits Cleavage Site(s)

Trypsin Serine lt 10 ng C-terminal sites of Arg and Lys

Pepsin Aspartic 15 ng Hydrophobic preferably aromatic residues in P1 and P1rsquo positions

Papain Cystine 15 ng Hydrolysis of proteins with broad specifi city for peptide bonds but preference for an amino acid bearing a large hydrophobic side chain at the P2 position Does not accept Val in P1rsquo

Carboxypeptidase A Metalloproteinase 50 ng Release of a C-terminal amino acid but little or no action with -Asp -Glu -Arg -Lys or -Pro

Components

Incubation buffer 25 mlFITC-casein solution 5 mlAssay buffer 200 mlProtease standard (trypsin) 20 μgFITC control 50 mgTrichloroacetic acid (TCA) solution 30 ml

PF0100-1KT 1 kit

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e D

etec

tio

n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

sigma-aldrichcom

LFL71112-508361

0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom

OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052

Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice

DevelopmentBulk Manufacturing Inquiries (800) 244-1173

Sigma-Aldrich3050 Spruce StreetSt Louis MO 63103 USAsigma-aldrichcom

Accelerating Customersrsquo

Success through Innovation and

Leadership in Life Science

High Technology and Service

ArgentinaSIGMA-ALDRICH DE ARGENTINA SA Free Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD Free Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

AustriaSIGMA-ALDRICH HANDELS GmbH Tel (+43) 1 605 81 10 Fax (+43) 1 605 81 20

BelgiumSIGMA-ALDRICH NVSAFree Tel 0800 14747 Free Fax 0800 14745 Tel (+32) 3 899 13 01 Fax (+32) 3 899 13 11

BrazilSIGMA-ALDRICH BRASIL LTDAFree Tel 0800 701 7425 Tel (+55) 11 3732 3100 Fax (+55) 11 5522 9895

CanadaSIGMA-ALDRICH CANADA LTDFree Tel 1800 565 1400 Free Fax 1800 265 3858 Tel (+1) 905 829 9500 Fax (+1) 905 829 9292

ChinaSIGMA-ALDRICH (SHANGHAI) TRADING CO LTDFree Tel 800 819 3336 Tel (+86) 21 6141 5566 Fax (+86) 21 6141 5567

Czech RepublicSIGMA-ALDRICH spol s r o Tel (+420) 246 003 200 Fax (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK AS Tel (+45) 43 56 59 10 Fax (+45) 43 56 59 05

FinlandSIGMA-ALDRICH FINLAND OYTel (+358) 9 350 9250 Fax (+358) 9 350 92555

FranceSIGMA-ALDRICH CHIMIE Sagraverl Free Tel 0800 211 408Free Fax 0800 031 052Tel (+33) 474 82 28 00 Fax (+33) 474 95 68 08

GermanySIGMA-ALDRICH CHEMIE GmbHFree Tel 0800 51 55 000 Free Fax 0800 64 90 000 Tel (+49) 89 6513 0 Fax (+49) 89 6513 1160

GreeceSIGMA-ALDRICH (OM) LTDTel (+30) 210 994 8010 Fax (+30) 210 994 3831

HungarySIGMA-ALDRICH Kft Ingyenes telefonszaacutem 06 80 355 355 Ingyenes fax szaacutem 06 80 344 344 Tel (+36) 1 235 9055 Fax (+36) 1 235 9050

IndiaSIGMA-ALDRICH CHEMICALS PRIVATE LIMITED Telephone Bangalore (+91) 80 6621 9600 New Delhi (+91) 11 4358 8000 Mumbai (+91) 22 2570 2364 Hyderabad (+91) 40 4015 5488 Fax Bangalore (+91) 80 6621 9650 New Delhi (+91) 11 4358 8001 Mumbai (+91) 22 2579 7589 Hyderabad (+91) 40 4015 5466

IrelandSIGMA-ALDRICH IRELAND LTD Free Tel 1800 200 888 Free Fax 1800 600 222Tel +353 (0) 402 20370Fax + 353 (0) 402 20375

IsraelSIGMA-ALDRICH ISRAEL LTD Free Tel 1 800 70 2222 Tel (+972) 8 948 4100 Fax (+972) 8 948 4200

ItalySIGMA-ALDRICH Srl Numero Verde 800 827018 Tel (+39) 02 3341 7310 Fax (+39) 02 3801 0737

JapanSIGMA-ALDRICH JAPAN KK Tel (+81) 3 5796 7300 Fax (+81) 3 5796 7315

KoreaSIGMA-ALDRICH KOREA Free Tel (+82) 80 023 7111 Free Fax (+82) 80 023 8111 Tel (+82) 31 329 9000 Fax (+82) 31 329 9090

MalaysiaSIGMA-ALDRICH (M) SDN BHDTel (+60) 3 5635 3321 Fax (+60) 3 5635 4116

MexicoSIGMA-ALDRICH QUIacuteMICA SA de CVFree Tel 01 800 007 5300 Free Fax 01 800 712 9920 Tel 52 722 276 1600 Fax 52 722 276 1601

The NetherlandsSIGMA-ALDRICH CHEMIE BVFree Tel 0800 022 9088 Free Fax 0800 022 9089 Tel (+31) 78 620 5411 Fax (+31) 78 620 5421

New ZealandSIGMA-ALDRICH NEW ZEALAND LTD Free Tel 0800 936 666 Free Fax 0800 937 777 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

NorwaySIGMA-ALDRICH NORWAY AS Tel (+47) 23 17 60 60 Fax (+47) 23 17 60 50

PolandSIGMA-ALDRICH Sp z oo Tel (+48) 61 829 01 00 Fax (+48) 61 829 01 20

PortugalSIGMA-ALDRICH QUIacuteMICA SAFree Tel 800 202 180 Free Fax 800 202 178 Tel (+351) 21 924 2555 Fax (+351) 21 924 2610

RussiaSIGMA-ALDRICH RUS LLC Tel +7 (495) 621 6037 +7 (495) 621 5828 Fax +7 (495) 621 5923

SingaporeSIGMA-ALDRICH PTE LTDTel (+65) 6779 1200 Fax (+65) 6779 1822

SlovakiaSIGMA-ALDRICH spol s r o Tel (+421) 255 571 562Fax (+421) 255 571 564

South AfricaSIGMA-ALDRICH SOUTH AFRICA (PTY) LTDFree Tel 0800 1100 75 Free Fax 0800 1100 79 Tel (+27) 11 979 1188 Fax (+27) 11 979 1119

SpainSIGMA-ALDRICH QUIacuteMICA SA Free Tel 900 101 376 Free Fax 900 102 028 Tel (+34) 91 661 99 77 Fax (+34) 91 661 96 42

SwedenSIGMA-ALDRICH SWEDEN ABTel (+46) 8 742 4200 Fax (+46) 8 742 4243

SwitzerlandSIGMA-ALDRICH CHEMIE GmbH Free Tel 0800 80 00 80 Free Fax 0800 80 00 81 Tel (+41) 81 755 2828 Fax (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTDFree Tel 0800 717 181 Free Fax 0800 378 785 Tel (+44) 1747 833 000 Fax (+44) 1747 833 313 SAFC (UK) Tel 01202 712305

United StatesSIGMA-ALDRICH PO Box 14508 St Louis Missouri 63178 Toll-Free 800 325 3010 Toll-Free Fax 800 325 5052 Call Collect (+1) 314 771 5750 Tel (+1) 314 771 5765 Fax (+1) 314 771 5757

VietnamSIGMA-ALDRICH PTE LTD VN ROTel (848) 3516 2810 Fax (848) 6258 4238

Internet sigma-aldrichcom

Page 30: Protease Inhibition and Detection

30 Order sigmacomorder Technical service sigmacomtechinfosigmacomlifescience

Pro

teas

e D

etec

tio

n

Non-Specific Protease Substrates

Casein from bovine milk[9000-71-9]

technical gradeC7078-500G 500 g

C7078-1KG 1 kg

C7078-10KG 10 kg

Casein NN-dimethyl ated from bovine milk

essentially salt-free lyophilized powderThis preparation has less than 10 reactivity with 246-trinitrobenzenesulfonic acid (TNBS) compared to non-methylated casein

Reductive methylation of C5890 by the method of Cabacungan JC et al Anal Biochem 124 272 (1982)

Protease substrate

C9801-1G 1 g

C9801-5G 5 g

C9801-25G 25 g

C9801-50G 50 g

Casein fluor escein iso thio cyanate from bovine milk

FITC-casein

highly sensitive protease substrate

Type II essentially salt-free lyophilized powderC3777-25MG 25 mg

C3777-100MG 100 mg

Type III essentially salt-free lyophilized powderC0528-10MG 10 mg

C0528-25MG 25 mg

C0528-100MG 100 mg

Elastin-orcein

Elastin impregnated with orcein

Prepared from E1625

Substrate for the estimation of elastase

E1500-5G 5 g

ElastinndashCongo red

Elastin impregnated with Congo red dye

Substrate for the estimation of elastase

E0502-5G 5 g

Gel atin from cold water fish skinTeleostean gel atin Gel atine [9000-70-8]Used as a blocking agent in immunochemistry

~45 in H2OG7765-250ML 250 mL

G7765-1L 1 L

Hemoglobin from bovine bloodBovine hemoglobin Methemoglobin Hb [9008-02-0]Oxygen transporter NO scavenger

suitable for protease substrate substrate powderPrepared from washed lysed and dialyzed erythrocytes

H2625-25G 25 g

H2625-100G 100 g

H2625-1KG 1 kg

HidendashRemazol Brilliant Blue RRemazol Brilliant Blue RndashHide Hide powder azure [37340-54-8]Hide powder covalently linked to Remazol Brilliant Blue R

Chromogenic substrate for trypsin

H6268-1G 1 g

H6268-5G 5 g

sigma-aldrichcom

Visit sigmacomenzymeexplorer

The New Enzyme Explorer

Redesigned homepage user interface

Expanded and reformatted technical resources and indexes

Protease Finder Identifies the cleavage sequence

Sigma and Sigma-Aldrich are registered trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LPThe red cube symbol is a trademark belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Ingenuity is a registered trademark belonging to Ingenuity Systems

Itrsquos the all-new YFG And itrsquos freeThink forward work smarter with Your Favorite Gene

powered by Ingenuity gt sigmacomyfg

Imagine a tool that gives you the power

to navigate a comprehensive universe

of continually curated biological data

and interactive genetic networks

all with a single simple click

sigma-aldrichcom

LFL71112-508361

0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom

OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052

Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice

DevelopmentBulk Manufacturing Inquiries (800) 244-1173

Sigma-Aldrich3050 Spruce StreetSt Louis MO 63103 USAsigma-aldrichcom

Accelerating Customersrsquo

Success through Innovation and

Leadership in Life Science

High Technology and Service

ArgentinaSIGMA-ALDRICH DE ARGENTINA SA Free Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD Free Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

AustriaSIGMA-ALDRICH HANDELS GmbH Tel (+43) 1 605 81 10 Fax (+43) 1 605 81 20

BelgiumSIGMA-ALDRICH NVSAFree Tel 0800 14747 Free Fax 0800 14745 Tel (+32) 3 899 13 01 Fax (+32) 3 899 13 11

BrazilSIGMA-ALDRICH BRASIL LTDAFree Tel 0800 701 7425 Tel (+55) 11 3732 3100 Fax (+55) 11 5522 9895

CanadaSIGMA-ALDRICH CANADA LTDFree Tel 1800 565 1400 Free Fax 1800 265 3858 Tel (+1) 905 829 9500 Fax (+1) 905 829 9292

ChinaSIGMA-ALDRICH (SHANGHAI) TRADING CO LTDFree Tel 800 819 3336 Tel (+86) 21 6141 5566 Fax (+86) 21 6141 5567

Czech RepublicSIGMA-ALDRICH spol s r o Tel (+420) 246 003 200 Fax (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK AS Tel (+45) 43 56 59 10 Fax (+45) 43 56 59 05

FinlandSIGMA-ALDRICH FINLAND OYTel (+358) 9 350 9250 Fax (+358) 9 350 92555

FranceSIGMA-ALDRICH CHIMIE Sagraverl Free Tel 0800 211 408Free Fax 0800 031 052Tel (+33) 474 82 28 00 Fax (+33) 474 95 68 08

GermanySIGMA-ALDRICH CHEMIE GmbHFree Tel 0800 51 55 000 Free Fax 0800 64 90 000 Tel (+49) 89 6513 0 Fax (+49) 89 6513 1160

GreeceSIGMA-ALDRICH (OM) LTDTel (+30) 210 994 8010 Fax (+30) 210 994 3831

HungarySIGMA-ALDRICH Kft Ingyenes telefonszaacutem 06 80 355 355 Ingyenes fax szaacutem 06 80 344 344 Tel (+36) 1 235 9055 Fax (+36) 1 235 9050

IndiaSIGMA-ALDRICH CHEMICALS PRIVATE LIMITED Telephone Bangalore (+91) 80 6621 9600 New Delhi (+91) 11 4358 8000 Mumbai (+91) 22 2570 2364 Hyderabad (+91) 40 4015 5488 Fax Bangalore (+91) 80 6621 9650 New Delhi (+91) 11 4358 8001 Mumbai (+91) 22 2579 7589 Hyderabad (+91) 40 4015 5466

IrelandSIGMA-ALDRICH IRELAND LTD Free Tel 1800 200 888 Free Fax 1800 600 222Tel +353 (0) 402 20370Fax + 353 (0) 402 20375

IsraelSIGMA-ALDRICH ISRAEL LTD Free Tel 1 800 70 2222 Tel (+972) 8 948 4100 Fax (+972) 8 948 4200

ItalySIGMA-ALDRICH Srl Numero Verde 800 827018 Tel (+39) 02 3341 7310 Fax (+39) 02 3801 0737

JapanSIGMA-ALDRICH JAPAN KK Tel (+81) 3 5796 7300 Fax (+81) 3 5796 7315

KoreaSIGMA-ALDRICH KOREA Free Tel (+82) 80 023 7111 Free Fax (+82) 80 023 8111 Tel (+82) 31 329 9000 Fax (+82) 31 329 9090

MalaysiaSIGMA-ALDRICH (M) SDN BHDTel (+60) 3 5635 3321 Fax (+60) 3 5635 4116

MexicoSIGMA-ALDRICH QUIacuteMICA SA de CVFree Tel 01 800 007 5300 Free Fax 01 800 712 9920 Tel 52 722 276 1600 Fax 52 722 276 1601

The NetherlandsSIGMA-ALDRICH CHEMIE BVFree Tel 0800 022 9088 Free Fax 0800 022 9089 Tel (+31) 78 620 5411 Fax (+31) 78 620 5421

New ZealandSIGMA-ALDRICH NEW ZEALAND LTD Free Tel 0800 936 666 Free Fax 0800 937 777 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

NorwaySIGMA-ALDRICH NORWAY AS Tel (+47) 23 17 60 60 Fax (+47) 23 17 60 50

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copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom

OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052

Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice

DevelopmentBulk Manufacturing Inquiries (800) 244-1173

Sigma-Aldrich3050 Spruce StreetSt Louis MO 63103 USAsigma-aldrichcom

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Success through Innovation and

Leadership in Life Science

High Technology and Service

ArgentinaSIGMA-ALDRICH DE ARGENTINA SA Free Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD Free Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

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PolandSIGMA-ALDRICH Sp z oo Tel (+48) 61 829 01 00 Fax (+48) 61 829 01 20

PortugalSIGMA-ALDRICH QUIacuteMICA SAFree Tel 800 202 180 Free Fax 800 202 178 Tel (+351) 21 924 2555 Fax (+351) 21 924 2610

RussiaSIGMA-ALDRICH RUS LLC Tel +7 (495) 621 6037 +7 (495) 621 5828 Fax +7 (495) 621 5923

SingaporeSIGMA-ALDRICH PTE LTDTel (+65) 6779 1200 Fax (+65) 6779 1822

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SpainSIGMA-ALDRICH QUIacuteMICA SA Free Tel 900 101 376 Free Fax 900 102 028 Tel (+34) 91 661 99 77 Fax (+34) 91 661 96 42

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SwitzerlandSIGMA-ALDRICH CHEMIE GmbH Free Tel 0800 80 00 80 Free Fax 0800 80 00 81 Tel (+41) 81 755 2828 Fax (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTDFree Tel 0800 717 181 Free Fax 0800 378 785 Tel (+44) 1747 833 000 Fax (+44) 1747 833 313 SAFC (UK) Tel 01202 712305

United StatesSIGMA-ALDRICH PO Box 14508 St Louis Missouri 63178 Toll-Free 800 325 3010 Toll-Free Fax 800 325 5052 Call Collect (+1) 314 771 5750 Tel (+1) 314 771 5765 Fax (+1) 314 771 5757

VietnamSIGMA-ALDRICH PTE LTD VN ROTel (848) 3516 2810 Fax (848) 6258 4238

Internet sigma-aldrichcom

Page 32: Protease Inhibition and Detection

LFL71112-508361

0039

copy2009 Sigma-Aldrich Co All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH FLUKA and SUPELCO are trademarks belonging to Sigma-Aldrich Co and its affiliate Sigma-Aldrich Biotechnology LP Sigma brand products are sold through Sigma-Aldrich Inc Sigma-Aldrich Inc warrants that its products conform to the information contained in this and other Sigma-Aldrich publications Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see reverse side of the invoice or packing slip Fused-Core is a trademark of Advanced Materials Technology Inc Chel-DE is a trademark of Ciba-Geigy AG Complexone is a registered trademark of CU Chemie Uetikon AG TWEEN is a registered trademark of Croda International PLC Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH Sepharose is a registered trademark of GE Healthcare IGEPAL is a registered trademark of General Dyestuff Corp Coomassie is a registered trademark of Imperial Chemical Industries Ltd TargeTron is a registered trademark of InGex LLC GenomePlex is a registered trademark of Rubicon Genomics Inc Prestige Antibodies ProteoSilver and SIGMAFAST are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co Ascentis and MISSION are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co UPLC is a registered trademark of Waters Corp

World Headquarters3050 Spruce St St Louis MO 63103(314) 771-5765sigma-aldrichcom

OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052

Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice

DevelopmentBulk Manufacturing Inquiries (800) 244-1173

Sigma-Aldrich3050 Spruce StreetSt Louis MO 63103 USAsigma-aldrichcom

Accelerating Customersrsquo

Success through Innovation and

Leadership in Life Science

High Technology and Service

ArgentinaSIGMA-ALDRICH DE ARGENTINA SA Free Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD Free Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

AustriaSIGMA-ALDRICH HANDELS GmbH Tel (+43) 1 605 81 10 Fax (+43) 1 605 81 20

BelgiumSIGMA-ALDRICH NVSAFree Tel 0800 14747 Free Fax 0800 14745 Tel (+32) 3 899 13 01 Fax (+32) 3 899 13 11

BrazilSIGMA-ALDRICH BRASIL LTDAFree Tel 0800 701 7425 Tel (+55) 11 3732 3100 Fax (+55) 11 5522 9895

CanadaSIGMA-ALDRICH CANADA LTDFree Tel 1800 565 1400 Free Fax 1800 265 3858 Tel (+1) 905 829 9500 Fax (+1) 905 829 9292

ChinaSIGMA-ALDRICH (SHANGHAI) TRADING CO LTDFree Tel 800 819 3336 Tel (+86) 21 6141 5566 Fax (+86) 21 6141 5567

Czech RepublicSIGMA-ALDRICH spol s r o Tel (+420) 246 003 200 Fax (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK AS Tel (+45) 43 56 59 10 Fax (+45) 43 56 59 05

FinlandSIGMA-ALDRICH FINLAND OYTel (+358) 9 350 9250 Fax (+358) 9 350 92555

FranceSIGMA-ALDRICH CHIMIE Sagraverl Free Tel 0800 211 408Free Fax 0800 031 052Tel (+33) 474 82 28 00 Fax (+33) 474 95 68 08

GermanySIGMA-ALDRICH CHEMIE GmbHFree Tel 0800 51 55 000 Free Fax 0800 64 90 000 Tel (+49) 89 6513 0 Fax (+49) 89 6513 1160

GreeceSIGMA-ALDRICH (OM) LTDTel (+30) 210 994 8010 Fax (+30) 210 994 3831

HungarySIGMA-ALDRICH Kft Ingyenes telefonszaacutem 06 80 355 355 Ingyenes fax szaacutem 06 80 344 344 Tel (+36) 1 235 9055 Fax (+36) 1 235 9050

IndiaSIGMA-ALDRICH CHEMICALS PRIVATE LIMITED Telephone Bangalore (+91) 80 6621 9600 New Delhi (+91) 11 4358 8000 Mumbai (+91) 22 2570 2364 Hyderabad (+91) 40 4015 5488 Fax Bangalore (+91) 80 6621 9650 New Delhi (+91) 11 4358 8001 Mumbai (+91) 22 2579 7589 Hyderabad (+91) 40 4015 5466

IrelandSIGMA-ALDRICH IRELAND LTD Free Tel 1800 200 888 Free Fax 1800 600 222Tel +353 (0) 402 20370Fax + 353 (0) 402 20375

IsraelSIGMA-ALDRICH ISRAEL LTD Free Tel 1 800 70 2222 Tel (+972) 8 948 4100 Fax (+972) 8 948 4200

ItalySIGMA-ALDRICH Srl Numero Verde 800 827018 Tel (+39) 02 3341 7310 Fax (+39) 02 3801 0737

JapanSIGMA-ALDRICH JAPAN KK Tel (+81) 3 5796 7300 Fax (+81) 3 5796 7315

KoreaSIGMA-ALDRICH KOREA Free Tel (+82) 80 023 7111 Free Fax (+82) 80 023 8111 Tel (+82) 31 329 9000 Fax (+82) 31 329 9090

MalaysiaSIGMA-ALDRICH (M) SDN BHDTel (+60) 3 5635 3321 Fax (+60) 3 5635 4116

MexicoSIGMA-ALDRICH QUIacuteMICA SA de CVFree Tel 01 800 007 5300 Free Fax 01 800 712 9920 Tel 52 722 276 1600 Fax 52 722 276 1601

The NetherlandsSIGMA-ALDRICH CHEMIE BVFree Tel 0800 022 9088 Free Fax 0800 022 9089 Tel (+31) 78 620 5411 Fax (+31) 78 620 5421

New ZealandSIGMA-ALDRICH NEW ZEALAND LTD Free Tel 0800 936 666 Free Fax 0800 937 777 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500

NorwaySIGMA-ALDRICH NORWAY AS Tel (+47) 23 17 60 60 Fax (+47) 23 17 60 50

PolandSIGMA-ALDRICH Sp z oo Tel (+48) 61 829 01 00 Fax (+48) 61 829 01 20

PortugalSIGMA-ALDRICH QUIacuteMICA SAFree Tel 800 202 180 Free Fax 800 202 178 Tel (+351) 21 924 2555 Fax (+351) 21 924 2610

RussiaSIGMA-ALDRICH RUS LLC Tel +7 (495) 621 6037 +7 (495) 621 5828 Fax +7 (495) 621 5923

SingaporeSIGMA-ALDRICH PTE LTDTel (+65) 6779 1200 Fax (+65) 6779 1822

SlovakiaSIGMA-ALDRICH spol s r o Tel (+421) 255 571 562Fax (+421) 255 571 564

South AfricaSIGMA-ALDRICH SOUTH AFRICA (PTY) LTDFree Tel 0800 1100 75 Free Fax 0800 1100 79 Tel (+27) 11 979 1188 Fax (+27) 11 979 1119

SpainSIGMA-ALDRICH QUIacuteMICA SA Free Tel 900 101 376 Free Fax 900 102 028 Tel (+34) 91 661 99 77 Fax (+34) 91 661 96 42

SwedenSIGMA-ALDRICH SWEDEN ABTel (+46) 8 742 4200 Fax (+46) 8 742 4243

SwitzerlandSIGMA-ALDRICH CHEMIE GmbH Free Tel 0800 80 00 80 Free Fax 0800 80 00 81 Tel (+41) 81 755 2828 Fax (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTDFree Tel 0800 717 181 Free Fax 0800 378 785 Tel (+44) 1747 833 000 Fax (+44) 1747 833 313 SAFC (UK) Tel 01202 712305

United StatesSIGMA-ALDRICH PO Box 14508 St Louis Missouri 63178 Toll-Free 800 325 3010 Toll-Free Fax 800 325 5052 Call Collect (+1) 314 771 5750 Tel (+1) 314 771 5765 Fax (+1) 314 771 5757

VietnamSIGMA-ALDRICH PTE LTD VN ROTel (848) 3516 2810 Fax (848) 6258 4238

Internet sigma-aldrichcom