& adp ribosylation highlighthps ank vit, vwa mvp-bd 1 444 88 191 259 nes mls introduction...

P R O D U C T F LY E R

highlight The PARP Family & ADP Ribosylation

CONTENTSPARP Family Antibodies 2PARP Family Proteins 2PARP Inhibitors 3PARP-10 & c-Myc 4XRCC1 Antibodies 5Inhibitors of Sirtuins [Class III HDACs] 5NAD+ Metabolism/NAD Kinases 6ARTs [ADP-ribosyltransferases] Antibodies 7P2X [Purinergic Receptors] Antibodies 7CD39 Family Antibodies 7Product Highlights 8

Tomorrow’s Reagents Manufactured Today® International Version

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Zn Finger, RRM

BRCT

Macro

NLS, NoLS

PARP domain

WGR

WWE, UlM

SAM

HPS

ANK

VIT, vWA

MVP-BD

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MLS

IntroductionPoly(ADP-ribosyl)ation is a post-translation-

al modification process which plays a critical role in diverse cellular functions such as DNA damage detection and repair, transcriptional regulation, intracellular trafficking, chromatin modification, mitotic apparatus formation, and apoptosis and necrosis. The transfer of ADP-ribose (ADPr) units onto protein acceptors is catalyzed by the family of poly(ADP-ribose) polymerases (PARPs), which members are en-coded in humans by 18 different genes [1] (see right side). PARP-1 is the founding member of the PARP family [2, 3].

The PARP Family

Picture courtesy of J.C. Amé, V. Schreiber and G. de Murcia (CNRS, Strasbourg) CONTINUED ON PAGE 2

DR2313ALX-270-452-M001 1 mg ALX-270-452-M005 5 mg ALX-270-452-M025 25 mg LIT: A newly synthesized poly(ADP-ribose) polymerase inhibitor, DR2313: pharmacological profiles, neuroprotective effects, and therapeutic time window in cerebral ischemia in rats: H. Naka-jima, et al.; J. Pharmacol. Exp. Ther. 312, 472 (2005)

3-Methyl-5-AIQ . HCl ALX-270-450-M001 1 mg ALX-270-450-M005 5 mg LIT: Synthesis and PARP-1 inhibitory activity of 3-substituted ana-logues of the potent water-soluble PARP inihbitor 5-aminoiso-quinolin-1-one (5-AIQ): E. C. Y. Woon, et al.; Bioorg. Med. Chem. (submitted), (2006)

For more information see Page 3!

The Widest Range of PARP Inhibitors!

DR2313

3-Methyl-5-AIQ . HCl

F LY E R N A M E E I N G E B E N

For updated prices and additional information visit www.alexis-biochemicals.com, contact your Local Distributor, or call +41 61 926 89 89

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Product Clone / Isotype Source / Host Specificity Application Prod. No. Size

MAb to Poly(ADP-ribose) [PAR] Clone: 10H / Mouse IgG3 Human, mouse, rat FC, ICC, IHC (PS), WB ALX-804-220-R100 100 µl

PAb to Poly(ADP-ribose) [PAR] Ab: 96-10-04 / From rabbit Human, mouse, rat ELISA, ICC, WB ALX-210-890-R100 100 µl

PAb to PARP-1 (mouse) From rabbit (supernatant) Mouse, human (weak) ICC, IHC, IP, WB ALX-210-619-R100 100 µl

PAb to PARP-1 (human) From rabbit (supernatant) Human, mouse (weak) ICC, IHC, IP, WB ALX-210-895-R100 100 µl

MAb to PARP-1 Clone: C2-10 / Mouse IgG1 Human, mouse, rat, primate ELISA, ICC, IP, WB ALX-804-210-R050 50 µl

MAb to PARP-1 Clone: F1-23 / Mouse IgG1 Human, bovine ELISA, ICC, IP, WB, FUNC ALX-804-211-R050 50 µl

PAb to PARP-1 From rabbit Human, mouse, bovine, chicken ELISA, ICC, IHC (FS, PS) ALX-210-302-R100 100 µl

PAb to PARP-1 From rabbit Human, mouse, rat, monkey, bovine ICC, IP, WB, FUNC ALX-210-221-R100 100 µl

PAb to PARP-1 (human) From goat (serum) Human, mouse WB ALX-210-897-R100 100 µl

PAb to PARP-1 (197-214) From rabbit Human, bovine, chicken ELISA, ICC, IP, WB ALX-210-219-R100 100 µl

PAb to PARP-1 (215-228) From rabbit Human, bovine, chicken ELISA, ICC, WB ALX-210-220-R100 100 µl

PAb to PARP-1 (509-524) From rabbit Human, mouse, rat, bovine ELISA, IP, WB, FUNC ALX-210-222-R100 100 µl

PAb to PARP-1 (BRCT Domain) From rabbit Human, mouse IP, WB ALX-210-540-R100 100 µl

MAb to PARP-2 Clone: 4G8 / Mouse IgG1 Human, mouse ELISA, WB ALX-804-639-L001 1 ml

PAb to PARP-2 Ab: Yuc / From rabbit Human, mouse, monkey ELISA, ICC, IP, WB ALX-210-303-R100 100 µl

PAb to PARP-2 Ab: Yuc / From rabbit, purified Human, mouse, monkey ICC ALX-210-896-R100 100 µl

PAb to PARP-2 (mouse) From rabbit (supernatant) Mouse, human (weak) ICC, IHC, IP, WB ALX-210-899-R100 100 µl

MAb to PARP-3 Clone: LA6B10 / Mouse IgM Human, mouse ICC ALX-804-466-R100 100 µl

PAb to PARP-3 (human) From rabbit Human IHC (PS), WB ASB-ARP33760-P050 50 µg

MAb to VPARP/PARP-4 Clone: p193-4 / Mouse IgG1 Human ICC, IHC (FS, PS), IP, WB ALX-801-023-C125 125 µg

MAb to VPARP/PARP-4 Clone: p193-6 / Mouse IgG2b Human ICC, IHC (FS), WB ALX-801-024-C125 125 µg

MAb to VPARP/PARP-4 Clone: p193-10 / Mouse IgG2a Human IHC, WB ALX-801-025-C125 125 µg

MAb to Tankyrase-1 [PARP-5a] Clone: 19A449 / Mouse IgG Human, mouse WB ALX-804-234-C100 100 µg

MAb to PARP-10 (human) Clone: 5H11 / Rat IgG1 Human ICC, IP, WB ALX-804-626-L001 1 ml

PAb to PARP-10 (300-350) (human) From rabbit Human IP, WB BET-A300-665A 0.1 mg

PAb to PARP-16 (human) From rabbit Human WB ASB-ARP33751-P050 50 µg

PAb to PARN (mouse) From rabbit Mouse WB ASB-ARP37393-T200 200 µg

DNA Repair (Cell Survival)PARP-1 has been implicated in multiple DNA

repair pathways including single-strand break (SSB) repair, double strand break (DSB) repair and base excision repair (BER). The molecular mechanism of BER may involve a local chro-matin relaxation mediated either by covalent modification of histones with poly(ADP-ribose) or by non-covalent interaction of histones with poly(ADP-ribose) that automodifies the enzyme. This might regulate the accessibility to other DNA repair proteins. In addition PARP-1 or poly(ADP-ribose) (PAR) may directly recruit other repair pro-teins like XRCC1 (X-ray repair cross-complement-ing 1) to the site of DNA damage [4-7]. PARP-2 is the only other known member of the PARP family to be activated by DNA strand breaks [8]. Although its DNA-binding domain is different from that of PARP-1, PARP-2 has been shown to be required for efficient BER through interactions with XRCC1 and PARP-1 [9].

BRCA1 and BRCA2 are tumor suppressor pro-teins important for DSB repair by homologous recombination. For a review see [13]. Recently it has been demonstrated that BRCA1 or BRCA2 deficient cells have lost their ability to repair sin-gle-strand breaks by homologous recombination after PARP-1 depletion or inhibition, which can result in cell cycle arrest and apoptosis [10, 11]. This specific killing of tumor cells may be of ther-apeutic importance for some human cancers.

PARP Family Proteins & Related ProductsProduct Source Purity Prod. No. Size

PARP-1 (human) (rec.) Produced in Sf9 cells ≥99% ALX-201-063-C020 20 µg

PARP-1 (human) (rec.) (His) Produced in Sf21 cells ≥98% ALX-201-250-C010 10 µg

PARP-1 (Cat. Dom.) (human) (rec.) Produced in Sf21 cells >90 % ALX-201-232-C020 20 µg

PARP-1 (BRCT Domain) (human) (rec.) (His) Produced in E. coli ≥98% ALX-201-255-C020 20 µg

PARP-1 (E988K mutant) (human) (rec.) Produced in Sf21 cells ≥95 % ALX-201-254-C010 10 µg

PARP-1b (human) (rec.) (active) (His) Produced in Sf21 cells ≥95% ALX-201-253-C020 20 µg

PARP-1 (bovine) Isolated from calf thymus ~90% ALX-202-042-C010 10 µg

PARP-1 (bovine) Isolated from calf thymus ~98% ALX-202-046-C010 10 µg

PARP-1 (bovine) (automod.) Standard Isolated from calf thymus ALX-202-044-R100 100 µl

PARP-2 (mouse) (rec.) Produced in Sf9 cells ≥98% ALX-201-064-C020 20 µg

PARP-2 (mouse) (rec.) (His) Produced in Sf21 cells ≥98% ALX-201-251-C010 10 µg

PARP-3 (human) (rec.) Produced in E. coli ≥97% ALX-201-170-C020 20 µg

PARP-3 (human) (rec.) (His) Produced in Sf21 cells ≥98% ALX-201-252-C010 10 µg

Poly(ADP-ribose) [PAR] Standard Automodified PARP-1 ≥95% ALX-202-043-C001 1 µg

PARG (bovine) Isolated from calf thymus ALX-202-045-UC01 0.1 U

PARP Family Antibodies & Related Products

CONTINUED ON PAGE 4

Latest InsightHormones trigger dramatic changes in the

structure and transcriptional activity of specific promoters that lead to exchange of repression complexes for activation complexes. Ju, et al. show that estrogen-dependent restructuring and transcription of the pS2 promoter require

the generation of a DNA double-strand break by a novel protein complex containing two en-zymes, topoisomerase IIβ and poly(ADP-ribose) polymerase (PARP).LIT: A topoisomerase IIbeta-mediated dsDNA break required for reg-ulated transcription: B.G. Ju, et al.; Science 312, 1798 (2006)

T H E PA R P FA M I LY

F LY E R N A M E E I N G E B E N

Purified (PF) = Purified (Preservative free); FC = Flow Cytometry; ICC = Immunocytochemistry; IP = Immunoprecipitation; IHC = Immunohistochemistry (FS = Frozen Sections, PS = Paraffin Sections); WB = Western blot; BP = Blocking Peptide

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Chemical Structures

The Widest Panel of PARP Inhibitors

PARP Inhibitors5-AIQ . HCl[5-Aminoisoquinolinone . HCl]ALX-270-285-M001 1 mgALX-270-285-M005 5 mg

3-Methyl-5-AIQ . HClALX-270-450-M001 1 mg ALX-270-450-M005 5 mgWater soluble, potent inhibitor of poly (ADP-ribose) polymerase-1 (PARP-1) (IC50=0.23µM) in vitro.

3-AminobenzamideALX-270-044-G001 1 gALX-270-044-G005 5 g

4-Amino-1,8-naphthalimideALX-270-250-M010 10 mg

BenzamideALX-270-174-G005 5 g

DAM-TIQ-A [UPF 796; 2-Dimethylaminomethyl-4H-thieno [2,3-c]iso-quinolin-5-one . HCl]ALX-270-431-M001 1 mg ALX-270-431-M005 5 mgWater soluble, mixed PARP-1/-2 inhibitor with excel-lent bioavailability and in vivo action.

DPQALX-270-221-M001 1 mg ALX-270-221-M005 5 mg

DR2313[2-Methyl-3,5,7,8-tetrahydrothiopyrano[4,3-d]pyrimidine-4-one]ALX-270-452-M001 1 mgALX-270-452-M005 5 mgALX-270-452-M025 25 mgPotent competitive PARP inhibitor (IC50=0.20µM and 0.24µM for PARP-1 and PARP-2 respectively).LIT: A newly synthesized poly(ADP-ribose) polymerase inhibitor, DR2313: pharmacological profiles, neuroprotective effects, and thera-peutic time window in cerebral ischemia in rats: H. Nakajima, et al.; J. Pharmacol. Exp. Ther. 312, 472 (2005)

EB-47 . 2HCl . 2H2OALX-270-383-M001 1 mg ALX-270-383-M005 5 mg

4-Hydroxyquinazoline [4-Quinazolinol]ALX-270-279-G001 1g

5-Iodo-6-amino-1,2-benzopyrone [INH2BP]ALX-270-278-M001 1 mg ALX-270-278-M005 5 mg

1,5-IsoquinolinediolALX-480-039-M005 5 mg ALX-480-039-M025 25 mg

NicotinamideALX-460-008-G010 10 g

NU1025 [8-Hydroxy-2-methylquinazoline-4-one]ALX-270-370-M001 1 mg ALX-270-370-M005 5 mg ALX-270-370-M025 25 mg

6(5H)-PhenanthridinoneALX-270-251-M010 10 mg

PJ-34ALX-270-289-M001 1 mg ALX-270-289-M005 5 mg

TIQ-A [Thieno[2,3-c]isoquinolin-5-one]ALX-270-365-M001 1 mg ALX-270-365-M005 5 mg

PARP ActivatorPeroxynitrite . tetramethylammoniumALX-400-036-L001 1 mlALX-400-036-5001 5x1 ml LIT: Peroxynitrite-induced cytotoxicity: mechanism and opportuni-ties for intervention: L. Virag, et al.; Toxicol. Lett. 140-141, 113 (2003) (Review)

5-AIQ . HCl 3-Aminobenzamide4-Amino-1,8-naphthalimide

BenzamideDPQ

EB-47 . 2HCl . 2H2O

4-Hydroxyquinazoline

5-Iodo-6-amino-1,2-benzopyrone

DAM-TIQ-A1,5-Isoquinolinediol

NicotinamidePeroxynitrite . tetramethyl-ammonium

TIQ-A NU1025

6(5H)-Phenanthridinone

PJ-34

Selected Latest Review Articles3. Life or death decisions: the cast of poly(ADP-ribose) poly-me ra se (PARP) as a therapeutic target for brain ischaemia: R. Pellicciari, et al.; Prog. Med. Chem. 42, 125 (2004) Poly(ADP-ribose) polymerase and the therapeutic effects of its inhibi-tors: P. Jagtap & C. Szabo; Nat. Rev. Drug Discov. 4, 421 (2005)

Targeting poly(ADP-ribosyl)ation: a promising approach in cancer therapy: J. F. Haince, et al.; Trends Mol. Med. 11, 456 (2005) Clinical perspectives of PARP inhibitors: G. Graziani & C. Szabo; Pharmacol. Res. 52, 109 (2005) PARP inhibitors for cancer therapy: N. J. Curtin; Expert. Rev. Mol. Med. 7, 1 (2005)

Chemopotentiation by PARP inhibitors in cancer therapy: L. Tentori & G. Graziani; Pharmacol. Res. 52, 25 (2005) Poly(ADP-ribose)polymerase inhibition - where now?: E.C. Woon & M.D. Threadgill; Curr. Med. Chem. 12, 2373 (2005) The expand-ing role of poly(ADP-ribose) metabolism: current challenges and new perspectives: J.P. Gagne, et al.; Curr. Opin. Cell Biol. 18, 145 (2006) Inhibition of poly(ADP-ribose) polymerase in cancer: E.R. Plummer; Curr. Opin. Pharmacol. 6, 364 (2006

Latest InsightC.C. Alano, et al. report that PARP-1 enzy-

matic activity is inhibited by tetracycline de-rivatives such as minocycline, which have pre-viously been shown to have neuroprotective and anti-inflammatory activity. Minocycline blocked neuronal NAD+ depletion as well as PARP-1 activation and they confirmed a di-rect competitive inhibition of PARP-1 by mi-nocycline (Ki=13.8nM) by using recombinant PARP-1 in a cell-free assay.

Minocycline . HClALX-380-109-M050 50 mg Tetracycline derivative with antimicrobial activity. Inhibitor of angiogenesis, apoptosis and PARP-1. Anti-inflammatory and neuroprotective.LIT: Minocycline inhibits poly(ADP-ribose) polymerase-1 at na-nomolar concentrations: C.C. Alano, et al.; PNAS 103, 9685 (2006)

3-Methyl-5-AIQ . HCl

DR2313

Minocycline . HCl



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Apoptosis/Necrosis (Cell Death)

Whereas activation of PARP-1 by mild geno-toxic stimuli may facilitate DNA repair and cell survival, irreparable DNA damage triggers apop-totic or necrotic cell death. In several patholog-ic situations that involve massive DNA damage, excessive activation of PARP-1 depletes cellular reserve of NAD and its precursor ATP, leading to cell death via energy failure. It recently has been shown that ROS-mediated DNA damage triggers activation of the PARP-1 and subsequent cell death. Although PARP-1 mediated cell death is thought to be necrotic, recent reports have shown that PARP-1 mediated cell death also has many features in common with apoptotic forms of cell death. It is suggested that translocation of apoptosis-inducing factor (AIF) from mitochon-dria to the nucleus is required for PARP-1 medi-ated and caspase-independent cell death [12].

PARP and TranscriptionPARP-1 may regulate transcription by at least

two different mechanisms. It can act as a modu-lator of the chromatin structure, but also func-tions as a component of enhancer/promoter regulatory complexes. PARP-1 can interact with the enhancers and promoters of genes by a) di-rect sequence-specific binding to enhancers, b) recruitment via DNA binding transcription fac-tors (e.g. NF-κB), and c) direct binding to DNA structures.LIT: 1 Poly(ADP-ribose): novel functions for an old molecule: V. Schreiber, et al.; Nat. Rev. Mol. Cell Biol. 7, 517 (2006) 2 cDNA sequence, protein structure, and chromosomal location of the human gene for poly(ADP-ribose) polymerase: B. W. Cherney, et al.; PNAS 84, 8370 (1987) 3 Hu-man nuclear NAD+ ADP-ribosyltransferase: localization of the gene on chromosome 1q41-q42 and expression of an active human enzyme in Escherichia coli: H. Herzog, et al.; PNAS 86, 3514 (1989) 4 XRCC1 is specifically associated with poly(ADP-ribose) polymerase and negatively regulates its activity following DNA damage: M. Masson, et al.; Mol. Cell Biol. 18, 3563 (1998) 5 Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins: J. M. Pleschke, et al.; J. Biol. Chem. 275, 40974 (2000) 6 A requirement for PARP-1 for the assembly or stability of XRCC1 nuclear foci at sites of oxidative DNA damage: S. F. El-Khamisy, et al.; Nucleic Acids Res. 31, 5526 (2003) 7 The role of poly(ADP-ribose) in the DNA damage signaling network: M. Malanga & F. R. Althaus; Bio-chem. Cell Biol. 83, 354 (2005) 8 PARP-2, A novel mammalian DNA damage-dependent poly(ADP-ribose) polymerase: J. C. Ame, et al.; J. Biol. Chem. 274, 17860 (1999) 9 Poly(ADP-ribose) polymerase-2 (PARP-2) is required for efficient base excision DNA repair in association with PARP-1 and XRCC1: V. Schreiber, et al.; J. Biol. Chem. 277, 23028 (2002) 10 Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase: H. E. Bryant, et al.; Nature 434, 913 (2005) 11 Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy: H. Farmer, et al.; Nature 434, 917 (2005) 12 Mediation of cell death by poly(ADP-ribose) polymerase-1: D. W. Koh, et al.; Pharmacol. Res. 52, 5 (2005) 13 Role of BRCA1 and BRCA2 as regulators of DNA repair, transcription, and cell cycle in response to DNA damage: K. Yoshida and Y. Miki; Cancer Sci. 95, 866 (2004)

Selected Latest Review ArticlesThe PARP superfamily: J. C. Ame, et al.; Bioessays 26, 882 (2004) Poly(ADP-ribosyl)ation by PARP-1: ‚PAR-laying‘ NAD+ into a nuclear signal: M.Y. Kim, et al.; Genes Dev. 19, 1951 (2005) Poly(ADP-ribose): A. Bürkle; FEBS J. 272, 4576 (2005) Poly(ADP-ribose) polymerases: managing genome stability: M. L. Meyer-Ficca, et al.; Int. J. Biochem. Cell Biol. 37, 920 (2005) Ageing and PARP: A. Bürkle, et al.; Phar-macol. Res. 52, 93 (2005) Poly(ADP-ribose) polymerase-1 medi-ated caspase-independent cell death after ischemia/reperfusion: S. J. van Wijk & G. J. Hageman; Free Radic. Biol. Med. 39, 81 (2005)

Regulation of poly(ADP-ribose) metabolism by poly(ADP-ribose) glycohydrolase: where and when?: M. E. Bonicalzi, et al.; Cell. Mol. Life Sci. 62, 739 (2005) Poly(ADP-ribosyl)ation regulation of life and death in the nervous system: D. W. Koh, et al.; Cell. Mol. Life Sci. 62, 760 (2005) Structure and function of poly(ADP-ribose) polymerase-1: role in oxidative stress-related pathologies: L. Virag; Curr. Vasc. Pharmacol. 3, 209 (2005) Mediation of cell death by poly(ADP-ribose) polymerase-1: D. W. Koh, et al.; Pharmacol. Res. 52, 5 (2005) The emerging role of poly(ADP-ribose) polymerase-1 in longevity: A. Bürkle, et al.; Int. J. Biochem. Cell Biol. 37, 1043 (2005) Targeting poly(ADP-ribosyl)ation: a promising approach in cancer therapy: J.F. Haince, et al.; Trends Mol. Med. 11, 456 (2005) Players in the PARP-1 cell-death pathway: JNK1 joins the cast: C.C. Alano & R.A. Swanson; TIBS 31, 309 (2006) Poly(ADP-ribose): nov-el functions for an old molecule: V. Schreiber, et al.; Nat. Rev. Mol. Cell Biol. 7, 517 (2006)

Human PARP-3 (hPARP-3) (540aa, with an approx. mass of 67kDa) is a core component of the centrosome and preferentially localized to the daughter centriole throughout the cell cy-cle. The N-terminal domain (54 aa) of hPARP-3 is responsible for its centrosomal localization.

An attractive hypothesis is that the presence of both PARP-1 and PARP-3 at the centrosome may link the DNA damage surveillance network to the mitotic fidelity checkpoint.LIT: PARP-3 localizes preferentially to the daughter centriole and in-terferes with the G1/S cell cycle progression: A. Augustin, et al.; J. Cell Sci. 116, 1551 (2003)

PARP-3

FIGURE: PARP-3 (red) preferentially localizes to the daughter centriole. The mother centriole (green) is immunostained with anti-acetylated α-tubulin in 3T3 cells. Picture courtesy of C. Spenle hauer & G. de Murcia (CNRS, Strasbourg).


M. Yu, et al. [1] identified and character-ized a novel 150kDa protein, designated PARP-10. PARP-10 can associate with the oncopro-tein c-Myc and possesses PARP activity, capa-ble of poly (ADP-ribosyl)ating itself and core histones but neither Myc nor Max. PARP-10 shuttles between the nuclear and cytoplas-mic compartments that is controlled at least in part by a leucine-rich nuclear export se-quence. Functionally PARP-10 inhibits trans-formation of primary cells (rat embryo fibro-blasts). PARP-10 might play a role in the con-trol of cell proliferation. To regulate its target genes, c-Myc recruits several different cofac-tors. K. Lesniewicz, et al. [2] suggest that PARP-10 may function as an additional cofactor that is recruited by c-Myc.

H.Y. Chou, et al. [3] suggested that cell cy-cle-dependent phosphorylation of PARP-10 by CDK2 plays crucial functions in cell prolif-eration.LIT: 1 PARP-10, a novel Myc-interacting protein with poly(ADP-ri-bose) polymerase activity, inhibits transformation: M. Yu, et al.; Onco-gene 24, 1982 (2005) 2 Overlap of the gene encoding the novel poly(ADP-ribose) polymerase Parp10 with the plectin 1 gene and common use of exon sequences: K. Lesniewicz, et al.; Genomics 86, 38 (2005) 3 CDK-dependent activation of poly(ADP-ribose) polymerase member 10 (PARP10): H.Y. Chou, et al.; J. Biol. Chem. 281, 15201 (2006)

PARP-10

FIGURE: Catalytic domain of PARP-10: Picture courtesy of B. Lüscher (University Aachen)

MAb to PARP-10 (human) (5H11)ALX-804-626-L001 1 ml CLONE: 5H11. ISOT YPE: Rat IgG1. IMMUNOGEN: GST-tagged human PARP-10 (aa 1-907). SPECIFICIT Y: Hu-man. APPLICATION: ICC, IP, WBLIT: M. Yu, et al.; Oncogene 24, 1982 (2005)

The proto-oncogene c-Myc is implicated in various physiological processes, like cell growth, proliferation, loss of differentiation and cell death. It also has been implicated in the loss of dysfunction of insulin-producing beta cells in diabetes. Recent studies in mice suggest that c-Myc has unexpected functions during both self-renewal and the differentia-tion of stem and early progenitor cells.

In an effort to identify novel c-Myc-interact-ing proteins, M. Yu, et al. (Oncogene 24, 1982 (2005)) recently discovered that PARP-10 can associate with the oncoprotein c-Myc.

c-Myc

MAb to c-Myc (human) (4H3)ALX-804-627-L001 1 ml CLONE: 4H3. ISOT YPE: Rat IgG2a. IMMUNOGEN: GST-tagged recombinant human c-Myc (aa 1-262) fu-sion protein. SPECIFICIT Y: Recognizes human c-Myc. APPLICATION: ICC, IP, WB.LIT: Identification and characterization of specific DNA-binding com-plexes containing members of the Myc/Max/Mad network of tran-scriptional regulators: A. Sommer, et al.; J. Biol. Chem. 273, 6632 (1998)

MAb to c-Myc (human) (6A10)ALX-804-632-L001 1 ml CLONE: 6A10. ISOT YPE: Rat IgG2a. IMMUNOGEN: GST-tagged recombinant human c-Myc (aa 1-262) fu-sion protein. SPECIFICIT Y: Recognizes human c-Myc. APPLICATION: ICC, IP, WB.LIT: See ALX-804-627


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XRCC1Human XRCC1 (X-ray repair cross-comple-

menting 1) is involved in base excision repair (BER) and single strand break repair (SSBR) and thought to act as a scaffolding protein for other repair factors. It has been shown to physical-ly interact with several enzymes known to be involved in the repair of SSBs, including DNA ligase IIIα, DNA polymerase β, APE1, polynu-cleotide kinase/phosphatase, poly(ADP-ribose) polymerases 1 and 2 (PARP-1 and 2) and 8-oxo-guanine DNA glycosylase (OGG1) [1-5].

LIT: 1 XRCC1 polypeptide interacts with DNA polymerase beta and possibly poly (ADP-ribose) polymerase, and DNA ligase III is a novel molecular ‚nick-sensor‘ in vitro: K. W. Caldecott, et al.; Nucleic Acids Res. 24, 4387 (1996) 2 Reconstitution of DNA base excision-repair with purified human proteins: interaction between DNA polymerase beta and the XRCC1 protein: Y. Kubota, et al.; EMBO J. 15, 6662 (1996)

3 Role of XRCC1 in the coordination and stimulation of oxidative DNA damage repair initiated by the DNA glycosylase hOGG1: S. Mars-in, et al.; J. Biol. Chem. 278, 44068 (2003) 4 XRCC1 stimulates human polynucleotide kinase activity at damaged DNA termini and acceler-ates DNA single-strand break repair: C. J. Whitehouse, et al.; Cell 104, 107 (2001) 5 Poly(ADP-ribose) polymerase-2 (PARP-2) is required for efficient base excision DNA repair in association with PARP-1 and XRCC1: V. Schreiber, et al.; J. Biol. Chem. 277, 23028 (2002)

Antibodies to XRCC1

Three general classes of deacetylases have been identified in eukaryotes: class I (HDACs 1, 2, 3 & 8), class II (HDACs 4, 5, 6, 7, 9 & 10), and class III HDACs (sirtuins).

Class III HDACs (SIRT1-7; Sirtuins) are the mammalian orthologs of the yeast Sir2 (Silent Information Regulator 2) protein from Sac-charomyces cerevisiae and form a structurally distinct class of NAD-dependent enzymes. Al-though there are seven mammalian sirtuins, deacetylase activity and substrate have only been reported for SIRT1, -2, -3 and -6.

SIRT1 has been shown to deacetylate sev-eral transcription factors, including the tumor suppressor protein p53, members of the FOXO family, HES-1 and HEY-2, PPARγ, MyoD, CTIP2, p300, PGC-1α, and NF-κB and has been linked to the regulation of aging. The implication of SIRT1 in a variety of processes including inflam-matory responses, cell defense, survival/cell death, muscle development, cell cycle regula-tion, axonal degeneration, DNA repair, and fatty acid metabolism has been described.

SIRT2 was shown to deacetylate α-tubulin and co-localizing with the cytoplasmic tubu-lin network, being involved in development. It also showed neuroprotective functions, con-trols insulin/IGF-1 levels in the cytoskeleton of Alzheimer disease and deacetylates p53 in the cytoplasm.

The role of sirtuins in controlling basic cellu-lar functions generated efforts in the develop-ment of small-molecule regulators that mod-ulate their cellular function. Compounds that target sirtuins may be useful in the treatment of many diseases.

Standard Inhibitor of hSIRT1 & hSIRT2

Sirtinol[2-[(2-Hydroxynaphthalen-1-yl-methylene)amino]-N-(1-phenethyl)benzamide]ALX-270-308-M001 1 mgALX-270-308-M005 5 mg Specific cell permeable inhibitor of the sirtuin family of NAD-dependent deacetylases (ySir2: IC50=48µM; hSIRT1: IC50=131µM; hSIRT2: IC50=58µM) with no ef-fect on human HDAC1. Reported to inhibit Sir2p trans-criptional silencing activity in vivo (IC50=25µM) and NAD-dependent histone deacetylase activity of puri-fied recombinant yeast Sir2p (IC50=70µM) and hSIRT2 (IC50=40µM) in vitro.LIT: Identification of a class of small molecule inhibitors of the sirtuin family of NAD-dependent deacetylases by phenotypic screening: C.M. Grozinger, et al.; J. Biol. Chem. 276, 38837 (2001) Design, synthesis, and biological evaluation of sirtinol analogues as class III histone/pro-tein deacetylase (Sirtuin) inhibitors: A. Mai, et al.; J. Med. Chem. 48, 7789 (2005) Sirt1 inhibitor, Sirtinol, induces senescence-like growth arrest with attenuated Ras-MAPK signaling in human cancer cells: H. Ota, et al.; Oncogene 25, 176 (2006)

NEW SIRT1 Inhibitor

6-Chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamideALX-270-437-M001 1 mg Potent cell permeable and metabolically stable specif-ic inhibitor of hSIRT1 (IC50=98nM in vivo / IC50=38nM in vitro; compared to hSIRT2: IC50=19µM and hSIRT3: IC50=48µM) with no effect on human HDACs class I and class II, nor NAD glycohydrolase (IC50>100µg). In-hibits the deacetylation of p53 (IC50=1µM).LIT: Discovery of indoles as potent and selective inhibitors of the deacetylase SIRT1: A.D. Napper, et al.; J. Med. Chem. 48, 8045 (2005)

Chemical Structures

O

NH2NH

Cl

Sirtinol6-Chloro-2,3,4,9-tetra-hydro-1H-carbazole-1-carboxamide

Product Source / Host Specificity Application Prod. No. Size

PAb to XRCC1 (human) From rabbit Human ELISA, ICC, IP, WB ALX-210-304-R050 50 µl

PAb to XRCC1 (human) From rabbit Human ICC, IP, WB ALX-210-539-R100 100 µl

PAb to XRCC1 (human) (exon 17) (BL316) From rabbit Human ICC, IHC, IP, WB BET-A300-065A 0.1 mg

PAb to XRCC1 (human) (phosph.) (pSer518/pThr519/pThr523) (BL603) From rabbit Human WB BET-A300-059A 0.1 mg

PAb to XRCC1 (human) (phosph.) (pSer461) (BL606) From rabbit Human WB BET-A300-147A 0.1 mg

PAb to XRCC1 (human) (phosph.) (pSer475) (BL608) From rabbit Human WB BET-A300-227A 0.1 mg

PAb to XRCC1 (human) (phosph.) (pSer485/pThr488) (BL610) From rabbit Human WB BET-A300-231A 0.1 mg

Selected Latest Review ArticlesXRCC1 keeps DNA from getting stranded: L.H. Thompson & M.G. West; Mutat. Res. 459, 1 (2000) XRCC1 and DNA strand break repair: K.W. Caldecott; DNA Repair (Amst) 2, 955 (2003) Mouse models of XRCC1 DNA repair polymorphisms and cancer: W.C. Ladiges; Oncogene 25, 1612 (2006)

For more HDAC Reagents please visit www.axxora.com!


More Information? Please visit

Sirtuins [Class III HDACs] – Fight against Aging!

Selected Latest Review ArticlesThe Sir 2 family of protein deacetylases: J.M. Denu; Curr. Opin. Chem. Biol. 9, 431 (2005) Sirtuins (histone deacetylases III) in the cellular response to DNA damage – facts and hypotheses: M. Kruszewski & I. Szumiel; DNA Repair (Amst) 4, 1306 (2005)

Neuronal protection by sirtuins in Alzheimer‘s disease: T.S. Anekonda & P.H. Reddy; J. Neurochem. 96, 305 (2006) Sirt1: a metabolic master switch that modulates lifespan: I.B. Leibiger & P.O. Berggren; Nat. Med. 12, 34 (2006)

Latest InsightSIRT6 promotes normal DNA repair. Absence of mammalian SIRT6 leads to genomic in-stability and aging-like phenotype.LIT: Genomic instability and aging-like phenotype in the ab-sence of mammalian SIRT6: R. Mostoslavsky, et al.; Cell 124, 315 (2006)


6

NAD+ Metabolism

NAD: New Insights on a Key Signalling Player!The pyridine nucleotides NAD and phosphor-ylated NAD (NADP) are major redox carrier in all organisms. Recent years have shown an in-crease of signalling and gene regulatory proc-esses where NAD+ or NADP are metabolised. NAD+ serves as substrate for protein modifi-cation including protein deacetylation, and mono- and poly-ADP-ribosylation, while both NAD+ and NADP represent precursors of intra-cellular calcium-mobilizing molecules. It is be-yond doubt that NADP-mediated signal trans-duction does not only regulate metabolic path-ways, but holds a key position in the control of fundamental cellular processes. NAD+ kinases phosphorylate NAD+ to NADP and have poten-tial roles in stress signalling and ROS genera-tion. Components such as nicotinamide mono-nucleotide adenylyltransferase-1 (NMNAT1), which is involved in the NAD+-salvage pathway, and PARG might also contribute locally to the fine-tuning and the persistence, respectively, of PAR signalling at DNA-damage sites.

ADP-RibosylationNuclear ADP-ribosylation reactions are criti-

cal for many physiological and pathophysiolog-ical outcomes, such as cellular differentiation and proliferation, genome integrity, carcino-genesis, cell survival and cell death, inflamma-tion, and neuronal functions.

An interesting feature is the potential cross talk of mono- and poly-ADP-ribosylation reac-tions and other NAD+-dependent reactions, either directly through trans-ADP-ribosyla-tion or indirectly through modulation of the NAD+ levels.

Cross talk of SIRT1 and poly-ADP-ribosyla-tion reactions may provide balance between cell survival and cell death, longevity, and se-nescence. Moreover, SIRTs and mono- or poly-ADP-ribosylation pathways may provide a uni-fied network for multicellular eukaryotes to deal with nutritional supply and environmen-tal stress.

Various studies suggested different molecu-lar mechanisms for how mono- and poly-ADP-ribosylation reactions are integrated in diverse physiological processes. Mono- and poly-ADP-ribosylation reactions may act on the level of signalling, modulation of chromatin structure, and epigenetic histone code.

Key Latest Review ArticleNuclear ADP-ribosylation reactions in mammalian cells: where are we today and where are we going?: P.O. Hassa, et al.; Micro-biol. Mol. Biol. Rev. 70, 789 (2006)

Selected Latest Review ArticlesNew functions of a long-known molecule. Emerging roles of NAD in cellular signaling: M. Ziegler; Eur. J. Biochem. 267, 1550 (2000) The new life of a centenarian: signalling functions of NAD(P): F. Berger, et al.; TIBS 29, 111 (2004) NAD - new roles in signalling and gene regulation in plants: L. Hunt, et al.; New Phytologist 163, 31 (2004) A vital link between energy and signal transduction: M. Ziegler; FEBS J. 272, 4561 (2005) NAD+ and NADH in cellular functions and cell death: W. Ying; Front. Biosci. 11, 3129 (2006)

Protein Modifications

NA

D+

Deg

rad

atio

n

Free ADP-ribose and Derivatives

ADP-ribose-transfer Reactions

NAD+

Cell Signalling

Cyclic ADP-ribose

ADP-ribosyl-cyclase / NAD+ Glycohydrolases

Calcium Mobilization

NAADP cADPRP cADPR

O-Acetyl-ADP-ribose Mono (ADP-ribose) Poly (ADP-ribose)

NADP NAD+ Kinase

ADP ATP

Nicotinamide

NMNPRPP

NPRTase

Nicotinamideriboside

ATP

Nrk1

NAMN

NMNAT NaMNAT

ATP ATP

NAD+ Synthase

GlnGlu NAAD

Nicotinic acid

NaPRTase

L-Tryptophan

NA

D+

Bio

syn

thes

isA

DP

-rib

ose

M

etab

olis

m

SIRT-Family (SIRTUINs)

Protein Deacetylation

Gene Silencing Immune Response

Bacterial Toxins

DNA Damage & Repair

Chromatin-structureModulation

Cell Division

PARG IsoformsARH FamilyNudix O-acetyl-ADP-ribose Hydrolase

Regulation of Calcium EntryProtein Glycation

Mono ADP-ribose transferases (ARTs)

PARP Family

Redox Reactions

Energy Metabolism

NAD +-using Enzymes

De novo synthesis

PRPP

ATP

NAD+ Dehydrogenase

NAD+ synthetic and catabolic pathways

NADP

FIGURE: Adapted from Poly(ADP-ribose): novel functions for an old molecule: V. Schreiber, et al.; Nat. Rev. Mol. Cell Biol. 7, 517 (2006)


NAD Kinase (full length) (active) (human) (rec.) (His) ALX-201-235-C050 50 µg Produced in E. coli. Human NADK is fused at the N-ter-minus to a His-tag. QUANTIT Y: Sufficient for the prepa-ration of >10µmol of NADP from NAD and ATP in 1ml of a 10mM solution. Free of any nucleotide convert-ing side-activities. LIT: Structural and functional characterization of human NAD kinase: F. Lerner, et al.; BBRC 288, 69 (2001)

NMNAT1 (human) (rec.) (His)ALX-201-237-C100 100 µg Produced in E. coli. Full length human NMNAT1 is fused at the N-terminus to a His-tag. APPLICATION: Well suited for the synthesis of NAD due to high spe-cific activity (10-20U/mg) and high substrate selec-tivity compared to NMNAT3 (human) (recombinant) (Prod. No. ALX-201-238).LIT: Molecular cloning, chromosomal localization, tissue mRNA levels, bacterial expression, and enzymatic properties of human NMN ade-nylyltransferase: M. Emanuelli, et al.; J. Biol. Chem. 276, 406 (2001) Characterization of recombinant human nicotinamide mononucle-otide adenylyl transferase (NMNAT), a nuclear enzyme essential for NAD synthesis: M. Schweiger, et al.; FEBS Lett. 492, 95 (2001) For a comprehensive bibliography please visit our website.

NAD Kinase (NT truncated) (active) (human) (rec.) (His) ALX-201-236-C050 50 µg Produced in E. coli. Human NADK (aa 64-446) is fused at the N-terminus to a His-tag. QUANTIT Y: Sufficient for the preparation of >10µmol of NADP from NAD and ATP in 1ml of a 10mM solution. Free of any nu-cleotide converting side-activities.LIT: Structural and functional characterization of human NAD kinase: F. Lerner, et al.; BBRC 288, 69 (2001)

NMNAT3 (human) (rec.) (His)ALX-201-238-C100 100 µg Produced in E. coli. Full length human NMNAT3 is fused at the N-terminus to a His-tag. APPLICATION: Well suited for the synthesis of NAD analogs due to lower substrate selectivity compared to NMNAT1 (hu-man) (recombinant) (Prod. No. ALX-201-237).LIT: Structural characterization of a human cytosolic NMN/NaMN adenylyltransferase and implication in human NAD biosynthesis: X. Zhang, et al.; J. Biol. Chem. 278, 13503 (2003) Subcellular compart-mentation and differential catalytic properties of the three human nicotinamide mononucleotide adenylyltransferase isoforms: F. Berger, et al.; J. Biol. Chem. 280, 36334 (2005)

Highly Pure & Fully Active NAD Kinase


7

Antibodies to ARTs / P2X Receptors / CD39 Family

Mono-ADP-ribosyltransferases (ARTs; mARTs)

Enzyme-modulated mono(ADP-ribosyl)ation was originally identified as the mechanism of ac-tion of several bacterial toxins [1]. The diphteria, cholera, pertussis and clostridia toxins are mono-ADP-ribosyltransferases (ARTs; mARTs; EC 2.4.2.31), known to cause various patholo-gies after their translocation into mammalian host cells. Recently, enzymes related to these toxins have been identified in eukaryotic cells. The best known of these ARTs are ecto-enzymes that are either glycosylphosphatidylinositol (GPI)-anchored or secretory. The mono(ADP-ribosyl)ation is a post-translational modifica-tion resulting in the transfer of ADP-ribose from NAD+ to various acceptor proteins. In contrast to intracellular poly(ADP-ribosyl)transferases (PARPs), the ARTs transfer a single ADP-ribose and are thus mono-ARTs (mARTs) [2].

Mammalian ARTs constitute a family of struc-turally related proteins expressed on the cell surface or secreted in the extracellular compart-ment. Five paralogs (ART1-5) have been cloned. Only four of them are expressed in human due to a defective ART2 gene, and six in the mouse as the result of ART2 gene duplication (ART2.1, ART2.2) [2]. ARTs are expressed in different tis-sues [3] and are involved in many physiological processes, such as myogenesis and innate as well as adaptive immunity. (ADP-ribosyl)ation activates the purinergic receptor P2X7, induc-ing a cascade of reactions, finally leading to ap-optosis [4].

LIT: 1 ADP-ribosylation: K. Ueda & O. Hayaishi; Annu. Rev. Biochem. 54, 73 (1985) 2 The family of toxin-related ecto-ADP-ribosyltrans-ferases in humans and the mouse: G. Glowacki, et al.; Protein Sci. 11, 1657 (2002) 3 Use of genetic immunization to raise antibodies rec-ognizing toxin-related cell surface ADP-ribosyltransferases in native conformation: F. Koch-Nolte, et al.; Cell Immunol. 236, 66 (2005) 4 NAD-induced T cell death: ADP-ribosylation of cell surface proteins by ART2 activates the cytolytic P2X7 purinoceptor: M. Seman, et al.; Immunity 19, 571 (2003)

P2X ReceptorsP2X receptors are a family of cation-per-

meable ligand gated ion channels that open in response to extracellular adenosine 5‘-tri-phosphate (ATP). They belong to the family of purinergic receptors. Seven genes coding for P2X subunits have been identified, and referred to as P2X1-P2X7. Each functional P2X receptor consists of three protein subunits, which are ar-ranged to form a gated, ion-permeable pore. ATP binds to the extracellular loop of the P2X receptor, and evokes a conformational change in the structure that results in the opening of the pore. This allows cations such as Na+ and Ca2+ to enter the cell, leading to depolarization of the cell membrane and the activation of vari-ous Ca2+-sensitive intracellular processes.

The CD39 FamilyCD39 was originally characterized as an

activation marker, identified in B cells, mono-cytes, and subsets of activated natural killer (NK) cells and T lymphocytes. CD39 is an ecto-nucleotidase, recently termed NTPDase-1. To-day 8 family members of NTPDases (NTPDase1-8; the CD39 family) are known. NTPDases are Ca2+- and Mg2+-dependent ecto-enzymes or ecto-nucleotidases that hydrolyze extracellu-lar nucleoside 5’-triphosphate and nucleoside 5’-diphosphate. By controlling the extracellular nucleotide levels NTPDases consequently have effects on P2 nucleotide receptor signalling. Thereby they play an important role in many cellular processes and have an impact on many physiological functions.

Selected Latest Review ArticlesNucleotides and cd39: principal modulatory players in hemo-stasis and thrombosis: H. Zimmermann; Nat. Med. 5, 987 (1999)

Purine signaling and potential new therapeutic approach: possible outcomes of NTPDase inhibition: F.P. Gendron, et al.; Curr. Drug Targets 3, 229 (2002) Ectonucleotidases of CD39 family modulate vascular inflammation and thrombosis in trans-plantation: S.C. Robson, et al.; Semin. Thromb. Hemost. 31, 217 (2005)

Selected Latest Review ArticlesMolecular physiology of P2X receptors: R.A. North; Physiol. Rev. 82, 1013 (2002) Molecular structure of P2X receptors: T.M. Egan, et al.; Curr. Top. Med. Chem. 4, 821 (2004) Molecular properties of ATP-gated P2X receptor ion channels: C. Vial, et al.; TIPS 25, 487 (2004) Overview of the P2 receptors: J.M. Bo-eynaems, et al.; Semin. Thromb. Hemost. 31, 139 (2005) P2X receptors as cell-surface ATP sensors in health and disease: B.S. Khakh & R.A. North; Nature 442, 527 (2006)

Product Clone / Isotype Source / Host Specificity Application Prod. No. Size

Antibodies to ARTs [mARTs]

MAb to ART1 [CD296] (human) GUGU1-A3/Rat IgG2b Human FC, ICC ALX-802-020-L001 1 ml

PAb to ART1 [CD296] (human) From rabbit Human FC, ICC ALX-215-037-R100 100 μl

MAb to ART1 [CD296] (mouse) NOGU1-A111/Rat IgG2a Mouse FC, ICC ALX-802-028-L001 1 ml

PAb to ART1 [CD296] (mouse) From rabbit Mouse FC, ICC ALX-215-038-R100 100 μl

MAb to ART2.1 (mouse) GUGU2-B54/Rat IgG2a Mouse FC, ICC ALX-802-021-L001 1 ml

MAb to ART2.2 (mouse) NIKA-102/Rat IgG2a Mouse FC, ICC ALX-802-022-L001 1 ml

PAb to ART2.2 (mouse) From rabbit Mouse FC, ICC, FUNC ALX-215-039-R100 100 μl

MAb to ART3 (human) GUGU3-A51/Rat IgG2a Human FC, ICC ALX-802-023-L001 1 ml

MAb to ART3 (mouse) GUGU3-A34/Rat IgG2a Mouse FC, ICC ALX-802-024-L001 1 ml

MAb to ART4 [CD297] (human) NONI-B4/Rat IgG2a Human FC, ICC ALX-802-025-L001 1 ml

PAb to ART4 [CD297] (human) From rabbit Human FC, ICC ALX-215-042-R100 100 μl

MAb to ART4 [CD297] (mouse) GUGU4-A53/Rat IgG2a Mouse FC, ICC ALX-802-026-L001 1 ml

Antibodies to P2X Receptors

PAb to Purinergic Receptor P2X1 (human) From rabbit Human FC, ICC, WB ALX-215-032-R100 100 μl

PAb to Purinergic Receptor P2X1 (mouse) From rabbit Mouse FC, ICC, IP ALX-215-031-R100 100 μl

PAb to Purinergic Receptor P2X4 (human) From rabbit Human FC, ICC, IP ALX-215-034-R100 100 μl


MAb to Purinergic Receptor P2X7 (mouse) HANO43/Rat IgG2b Mouse FC, ICC, IP ALX-802-027-L001 1 ml


PAb to Purinergic Receptor P2X7 (rat) From rabbit Rat FC, ICC, IP ALX-215-036-R100 100 μl

Antibodies to CD39 Family

PAb to CD39 (mouse) From rabbit Mouse FC, ICC ALX-215-043-R100 100 μl

MAb to CD39L1 (mouse) NOGU3-B28/Rat IgG2b Mouse FC, ICC ALX-802-029-L001 1 ml

PAb to CD39L1 (human) From rabbit Human FC, ICC ALX-215-045-R100 100 μl

PAb to CD39L1 (mouse) From rabbit Mouse FC, ICC ALX-215-044-R100 100 μl

PAb to CD39L2 (mouse) From rabbit Mouse FC, ICC ALX-215-046-R100 100 μl

PAb to CD39L4 (human) From rabbit Human FC, ICC, IP ALX-215-047-R100 100 μl

PAb to CD39L4 (mouse) From rabbit Mouse FC, ICC, IP ALX-215-048-R100 100 μl


Selected Latest Review ArticlesFunctional aspects of protein mono-ADP-ribosylation: D. Corda & M. Di Girolamo; EMBO J. 22, 1953 (2003) Ecto-ADP-ribosyltrans-ferases (ARTs): emerging actors in cell communication and sign-aling: M. Seman, et al.; Curr. Med. Chem. 11, 857 (2004) Physi-ological relevance of the endogenous mono(ADP-ribosyl)ation of cellular proteins: M. Di Girolamo, et al.; FEBS J. 272, 4565 (2005)

ADP-ribosylation of membrane proteins: unveiling the secrets of a crucial regulatory mechanism in mammalian cells: F. Koch-Nolte, et al.; Ann. Med. 38, 188 (2006)


8

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01-N

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6

DNA Damage Marker

FIGURE: Detection of DNA damage induced globally (X-ray irradia-tion with 10 Gray) or locally (laser microbeam irradiation) with anti-γ-H2AX PAb (Prod. No. ALX-210-390). Picture courtesy of C. Spenle-hauer & G. de Murcia (CNRS, Strasbourg).

- + - +

3T3 cells

20Gy

γ-H2AX

Actin

Western blot

0 Gy 10 Gy

laser microbeam

FIGURE: Detection of DNA damage in HeLa cells induced locally with anti-γ-H2AX PAb (Prod. No. ALX-210-390). Right) merged im-age of anti-γ-H2AX PAb and HOE 33258 (Prod. No. ALX-620-051) staining. Pictures courtesy of C. Spenlehauer & G. de Murcia (CNRS, Strasbourg).

PAb to Histone γ-H2AX (human) (phosphorylated) (pSer139)ALX-210-390-R050 50 µl From rabbit. IMMUNOGEN: Synthetic peptide corre-sponding to Ser139-phosphorylated human γ-his-tone H2AX (K134ATQApSQEY142). SPECIFICIT Y: Recog-nizes Ser139-phosphorylated human H2AX. Does not cross-react with non-phosphorylated human H2AX. APPLICATION: ICC, WB.LIT: Local DNA damage by proton microbeam irradiation induces poly(ADP-ribose) synthesis in mammalian cells: L. Tartier, et al.; Mu-tagenesis 18, 411 (2003)

Clone 10H – The Standard DNA Damage Detection Marker

MAb to Poly(ADP-ribose) [PAR] (10H)ALX-804-220-R100 100 µl CLONE: 10H. ISOT YPE: Mouse IgG3. IMMUNOGEN: Pu-rified poly(ADP-ribose). SPECIFICIT Y: Recognizes poly(ADP-ribose) synthesized by a broad range of PARPs (poly(ADP-ribose) polymerases) like human, mouse, rat or Drosophila PARP enzyme. APPLICATION: FC, ICC, IHC (PS), WB. LIT: Monoclonal antibodies to poly(adenosine diphosphate ribose) recognize different structures: H. Kawamitsu, et al.; Biochemistry 23, 3771 (1984), (Original Reference) Detection of poly(ADP-ribose) polymerase and its reaction product poly(ADP-ribose) by immuno-cytochemistry: J.H. Küpper, et al.; Histochem. J. 28, 391 (1996) Mul-tiparametric staining to identify apoptotic human cells: C. Negri, et al.; Exp. Cell Res. 234, 174 (1997) Poly(ADP-ribose) immunostaining to detect apoptosis induced by a neurotoxic fragment of prion pro-tein: A. Bürkle, et al.; Histochem. J. 31, 711 (1999) Reactive oxygen species participate in mdr1b mRNA and P-glycoprotein overexpres-sion in primary rat hepatocyte cultures: C. Ziemann, et al.; Carcino-genesis 20, 407 (1999) Detection of poly(ADP-ribose) synthesis in Drosophila testes upon gamma-irradiation: S. Lankenau, et al.; Chro-mosoma 108, 44 (1999) Quantitative nonisotopic immuno-dot-blot method for the assessment of cellular poly(ADP-ribosyl)ation capacity: R. Pfeiffer, et al.; Anal. Biochem. 275, 118 (1999) Poly(ADP-ribosyl)ation, genomic instability, and longevity: A. Bürkle; Ann. N Y Acad. Sci. 908, 126 (2000) Flow-cytometric assessment of cellular poly(ADP-ribosyl)ation capacity in peripheral blood lymphocytes: A. Kunzmann, et al.; Immun. Ageing 3, 8 (2006)

FIGURE: HeLa irradiated cells with a microbeam laser. Picture courtesy of C. Spenlehauer & G. de Murcia (CNRS, Strasbourg)

FIGURE: Detection of DNA damage.

FIGURE: Detection of apoptotic cells by immunofluorescence.

Latest InsightCell survival after genotoxic stress is deter-

mined by a counterbalance of pro- and anti-death factors. Sirtuins (SIRTs) are deacetylases that promote cell survival whereas poly(ADP-ribose) polymerases (PARPs) can act both as survival and death inducing factor and the two protein families are strictly dependent on NAD+ for their activities. U. Kolthur-Seethar-am, et al. report that the functional interplay of SIRT1 and PARP-1 in response to DNA dam-age results in AIF (apoptosis inducing factor)-mediated cell death. These findings establish a functional link between the two NAD+-de-pendent enzyme systems and provide a phys-iological interpretation for the mechanism of death in cells lacking SIRT1.LIT: Control of AIF-mediated cell death by the functional inter-play of SIRT1 and PARP-1 in response to DNA damage: U. Kolthur-Seetharam, et al.; Cell Cycle 5, 873 (2006)

Bacterial Pertussis Toxin

Pertussis Toxin ALX-630-003-C050 50 µg Isolated from Bordetella pertussis. Pertussis toxin is the major protein toxin produced by virulent strains of Bordetella pertussis. The purified pro-tein consists of five dissimilar subunits: S-1 (MW 28kDa), S-2 (MW 23kDa), S-3 (MW 22kDa), S-4 (MW 11.7kDa) and S-5 (MW 9.3kDa), in a molar ratio of 1:1:1:2:1. S-1 (A protomer) is responsible for the enzymatic activity of the toxin. Together, S-2, S-3, S-4 and S-5 comprise the B oligomer, responsible for binding the toxin to the cell surface.LIT: Subunit structure of islet-activating protein, pertussis toxin, in conformity with the A-B model: M. Tamura, et al.; Biochemistry 21, 5516 (1982) Structure-activity analysis of the activation of pertus-sis toxin: H.R. Kaslow, et al.; Biochemistry 26, 123 (1987)

More Information? Please visit


MAb 10H DAPI Staining

H2O2 (1mM) Incubation

Control

MAb 10H DAPI Staining

ApoptosisInduction

Control

& adp ribosylation highlighthps ank vit, vwa mvp-bd 1 444 88 191 259 nes mls introduction...

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