Tools and Capabilities Focus on cell based assays

and gene expression analysis

for early phase Drug Discovery

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Focus:

Provide researchers a subset of tools and methods to enable

successful outcomes in early phase drug discovery. Failure is

success when determined early.

Value (simplified):

Physiologically relevant primary human (STEMEZ™), rat and

mouse cells Better in vitro screening of targets and molecules

Reduced animal model use

Shorten development timeline

Lower downstream costs

Images: STEMEZ™ hNP1™ can be

differentiated into various neuronal

subtypes under defined conditions.

DAPIGFAP Antibody

• > 80% Tuj1+

• > 60% MAP2+

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Why Neuromics? Standing out in a crowded room

We have a experience serving many large Pharmas, Biotechs,

Academic and Government Labs.

Customer success focused

Expert technical support

Detailed protocols and methods

Quick replacements and reorders

Large expert resource network

Published expert input

Customer feedback and data sharing through

social media outlets and blogs

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Testimonial: got 10 million cells total after extraction from the tissue. At Day 4 they all developed long axons.

Thank you so much for the replacement." Dr. Lidia Gardner, University of Tennessee HSC..

Gene Expression

Analysis

Media/3-D

Gels/ECMS

Apoptosis

Detection

Cell Migration

Assays

Growth

Factors/Markers Applications include:

•Proliferation and

neurogenesis

•Neuronal differentiation

•Neurite outgrowth

•Cell migration

•Neurotoxicity screening

•Cell based compound

screening

•G-protein coupled receptors

•Ion channels

•Stem cell transplantation

Derived and

manufactured

neural cells from

human

embryonic stem

cell line H9

(WA09).

Stable, Potent and Cost Effective Cells

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Markers/Antibodies Growth Factors/Culture Media Gene Expression Analysis Apoptosis Detection

Imges: GFAP (Catalog#:

aCH22102) Staining of of

Astrocytes in the Ventral

horn. Courtesy of Dr. Qi-lin

L. Cao, University of

Louisville School of

Medicine. Rat spinal cord

homogenate WBs: NF-H,

NF-M,, NF-L,, NF66 and

GFAP.

STEMEZTMhNP1

Human Neural

Progenitors treated

with hGDNF

cultured in

STEMEZ hNP1

Neural Progenitor

Expansion Media

i-Fect knockdown of Delta Opioid

Receptor. Molecular Pain 2005,

1:29

Jurkat cells dually

stained with Hoechst

and Polycaspase

Assay Kit,

Assay kits, Qualified Media and Reagents-Sample

Images

Cell Based Assays-Examples

STEMEZ hN2 Human Neural Discovery Kit

Phenotyping

Electrophysical and FLIPR Data Provided upon request

Neural phenotypes derived from N2 cell

lines. (A) Phase contrast image of

differentiated culture. (B) Network

including post-mitotic motoneurons

(HB9). (C) Cholinergic neuron. (D) Tuj-1

positive cells that are DAT-positive

(dopamine transporter; closed arrow)

and DAT-negative (open arrow). (E)

Gabaergic neurons, inset illustrates

GABA in axon, but not the dendrites

(arrow).

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Tools and Capabilities

Cell Based Assays-Examples

STEMEZ™ hNP1™ Human Neural Progenitor Kit is an efficient tool to quantitatively

detect both stimulators and inhibitors of cell migration when used with the OrisTM Cell

Migration Assay Kit (Platypus Technologies). Method development to date indicates that

the assay has the potential for adaptation as a homogenous HTS-suitable cell-based

assay.

Cell Based Assays-Examples

Primary Rat Neurons and Astrocytes

Transfection of functional HeV

glycoproteins and infection with HeV

pseudotyped virions.In order to establish

the feasibility of carrying out the proposed

experiments in primary neurons, we show

(figure ) that our assays are amenable to

use in primary neurons. In the experiment, Combined Hippocampus, Cortex, and Ventricular -E18

(Neuromics) were plated, and at 3 days were transfected with plasmids encoding HeV G/F as well

as YFP. On the following day, these cells were infected with HeV or VSV pseudotyped viruses

bearing RFP. In the figure, (A) the phase contrast photos show the differentiated neurons; (B)

upon excitation for RFP, the red fluorescence indicates neurons infected by HeV pseudotyped

virions; (C) upon excitation for YFP, and the green fluorescence shows the efficiency of

transfection in neurons. This experiment indicates that the proposed experiments can be carried

out in primary neurons, which are transfectable and infectable in our systems, and thus supports

all the proposed aims. Data Courtesy of Dr. Matteo Porotto, Weill Cornell Medical College. Larger

Image

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Cell Based Assays-Examples

Primary Rat Neurons and Astrocytes

Data Courtesy of Evanna Gleason, Louisiana State University

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Gene Expression Analysis It’s all about delivery!

i-FectTM siRNA delivery kit : Success in delivering siRNA to

DOR, hTERT, The β3 subunit of the Na+,K+-ATPase,

rSNSR1, NTS1. NAV1.8, Survivin, TRL4, Flaviviruses and

more.

Tools for delivering plasmids, proteins, antisense and

other small RNA and DNA molecule.

Transfection Delivery: Customer Publications

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More Key Product Category Links:

Antibodies-Markers

Apoptosis Research Reagents -includes:

detection kits, antibodies and proteins

Proteins Recombinant and natural proteins-

includes a wide selection of cell growth factors.

Transfection Reagents-Gene Expression Analysis

Cell Growth Media

Clear Understanding of where Neuromics could have

a positive impact. More intense in vitro screening of compounds and

targets.

Better candidates for downstream processes.

Propose solutions based on this understanding

Follow up, tweak and improve

3/5/2011

Pete Shuster

CEO and Owner

Direct Phone: 612-801-1007

pshuster@neuromics.com

Social Networking:

www.neuromics.com

www.neuromics.net

neuromics.blogspot.com

sirnatransfection.blogspot.com

3/5/2011