EMD Millipore is a division of Merck KGaA, Darmstadt, Germany

Small Molecule Libraries and PanelsNavigate more cellular pathways with chemical biology

2

Platforms, Technologies, and ServicesAs a tools provider and partner in research, EMD Millipore is committed to the advancement of life science research and therapeutic development. This guide outlines EMD Millipore’s portfolio of small molecule libraries and pathway-focused panels, which are designed to facilitate target identification, pathway detection and profiling. These reagents provide proven solutions for a range of applications, platforms and technologies, and are backed by extensive technical support.

CALBIOCHEM® SMALL MOLECULESSmall-molecule compounds, including inhibitors,

activators, and other pathway modulators, are critical

tools for researchers studying cell signaling and other

mechanisms that control cell fate, function and phenotype.

EMD Millipore’s Calbiochem® reagents have been cited

in thousands of peer-reviewed publications. From

libraries and pathway panels to individual reagents, the

Calbiochem® line of products offers the widest and most

cited selection of inhibitors and activators worldwide.

CELLS AND CELL CULTUREEMD Millipore’s innovative cell culture solutions help

optimize cell growth and maintenance. We offer an

extensive range of human and rodent stem cells, primary

cells and media designed for most types of stem cells,

including embryonic, mesenchymal, and neural stem

cells. These optimized media include serum-free, feeder-

free formulations, and are supported by our feeder cells,

supplements, reagents, and cultureware. Our flexible sterile

filtration devices offer fast flow and have many membrane

options. Also available are membrane-based cultureware to

mimic in vivo conditions and provide coculture options.

ANTIBODIES AND IMMUNOASSAYSWith the expertise of Upstate® and Chemicon®, EMD

Millipore provides an extensive, focused, validated portfolio

of antibodies and immunoassays, with breadth and depth

in major research areas backed by excellent service and

support. EMD Millipore also offers a variety of ELISAs in

major research areas, including cell health.

CELL-BASED ASSAYS AND QUANTITATIVE CELL IMAGINGOur portfolio of live cell, whole-cell and cell-based activity

assays and reporter systems advances direct and indirect

detection of biological processes. These technologies

facilitate protein target validation, identify cellular

pathways and determine mechanism of action for lead

optimization environments. EMD Millipore also offers

assays for high-content, multiparametric cell imaging,

enabling identification of cellular responses and events

under user-defined conditions.

FLOW CYTOMETRY ASSAYS AND SYSTEMSFlow cytometry is essential for in-depth cell analysis,

with the capacity to simultaneously measure multiple

parameters on individual cells. Our easyCyte™ flow

cytometers provide direct, precise measurement via

microcapillary technology that translates into smaller

samples, less reagents, and minimal waste. Our validated

FlowCellect™ assay kits and Milli-Mark™ conjugated

antibodies, along with application-specific analysis

software modules, provide a complete solution for flow

cytometry.

MILLIPLEX® map MULTIPLEX ASSAYSMILLIPLEX® map assays offer the broadest selection of

multiplex kits and reagents in a wide variety of research

areas, measuring multiple biomarkers using a small sample

size. MILLIPLEX® map enables the simultaneous detection

of multiple soluble or intracellular biomarkers. Using the

Luminex® xMAP® bead-based technology, these flexible

and customizable assays are exhaustively tested and

qualified for sensitivity, specificity, reproducibility and wide

dynamic range.

MOLECULAR BIOLOGY TOOLSFor every step of the molecular biology and protein

workflow, from cloning DNA targets to purifying native

recombinant proteins, EMD Millipore provides reagents,

kits, cells and tools that are specifically designed to meet

your scientific and technical goals, including the Novagen®

line of products for DNA amplification, purification, and

propagation and reagents for efficient transfection.

3

Platforms, Technologies, and Services

Why use small molecules to study biological processes?

Table of Contents

Introduction

Chemical biology has been used successfully as both gain-

of-function and loss-of-function approaches to study a

variety of biological processes. For example, in chemical

genetics, either small organic molecules or peptides are used

to activate or inhibit specific proteins/enzymes involved

in specific signal transduction pathways. This allows

researchers to analyze the phenotype when a specific cellular

protein is induced or suppressed.

Compared to other approaches such as over-expressing

plasmid DNA or genetic knockdowns, respectively, chemical

genetics is technically simple to perform in cell culture, and

requires fewer resources and less time. Compared to the

RNAi approach, small molecules provide the advantage of

acting quickly, and the effects can often be reversed rapidly

by simply washing1. The fast action of most small molecules

also makes them ideal for live imaging. In addition, the use

of small molecules can provide dose response information.

Small molecules offer a powerful approach to temporally

and spatially modulate individual proteins and processes

that can control biological phenotypes. Small molecules

have, for instance, helped to advance our understanding of

the biological pathways that influence stem cell fate, and

have also been used successfully to modulate self-renewal,

survival, and direct reprogramming and differentiation of

pluripotent stem cells. Further, small molecules have helped

to identify signaling molecules that define and maintain the

extensive intracellular communication networks that control

growth, differentiation, metabolism, and other critical

cellular functions.

Reference:1. Eggert, U.S. et al. (2006) Small molecules in an RNAi world.

Mol.BioSyst. 2, 93-96.

PG. 04

PG. 09

PG. 13

Analysis of cellular phenotype

Modulation of cell fate

Analysis of specific signaling pathways

InhibitorSelect™ Kinase Inhibitor Libraries

StemSelect® Library

Signaling Pathway Panels:

PI 3-K/Akt/mTOR EGFR JAK/STAT NF-kB MAP Kinase VEGF Wnt IGFFGF Hedgehog

4

Cellular phenotypes are complex phenomena that require

careful coordination of many cellular processes. In

examining underlying molecular mechanisms, small

molecules offer a powerful tool to identify several classes

of signaling molecules involved in these processes. Protein

kinase is one class of molecules that has been revealed

to control many cellular processes. Protein kinases have

been shown to mediate most of the signal transduction in

eukaryotic cells1. Protein kinases also control many other

cellular events such as metabolism, cell cycle progression,

apoptosis, and differentiation.

Reference1. Manning, G. et al. (2002) The protein kinase complement of the

human genome. Science 298, 1912-1934.

InhibitorSelect™ Kinase Inhibitor Libraries Recognizing both the tremendous opportunities and

critical role of protein kinases, we have introduced several

Calbiochem® InhibitorSelect™ Libraries that contain

collections of carefully selected, structurally diverse, and

potent small molecules targeting members from several

families of protein kinases in convenient, cost effective

formats.

Well-characterized: Unlike small molecule libraries from

other leading suppliers, InhibitorSelect™ libraries are

provided with documented cell permeability, reversibility,

potency, published IC50/Ki values, lot-specific data, stability,

and HPLC purity for most of the included molecules.

Well-documented: Comprehensive documentation about

each inhibitor, such as molecular structure, IC50 values,

literature citations, and CAS number (where available), is

provided at your fingertips

Visit www.emdbiosciences.com/inhibitorselect to learn more information about InhibitorSelect™ Libraries

Analysis of cellular phenotype

I.

Akt Inhibitor V, Triciribine (Cat. No. 124012)

Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 (Cat. No. 124018)

Lot Specific HPLC DataStructure Structure

Cell-Permeable YesReversible YesPurity 98.9%

1601501401301201101009080706050403020100

1 2 3 4 5 6 7 8 9 10

Cell-Permeable YesReversible YesPurity 97.6%

Lot Specific HPLC Data1601501401301201101009080706050403020100

1 2 3 4 5 6 7 8 9 10

H2N

CH3N

N

NNO

HO

OH OH

N

CH2

NH

NN

NO

NHN

N

Akt Inhibitor V, Triciribine (Cat. No. 124012)

Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 (Cat. No. 124018)

Lot Specific HPLC DataStructure Structure

Cell-Permeable YesReversible YesPurity 98.9%

1601501401301201101009080706050403020100

1 2 3 4 5 6 7 8 9 10

Cell-Permeable YesReversible YesPurity 97.6%

Lot Specific HPLC Data1601501401301201101009080706050403020100

1 2 3 4 5 6 7 8 9 10

H2N

CH3N

N

NNO

HO

OH OH

N

CH2

NH

NN

NO

NHN

N

Two Akt inhibitors included in EMD Millipore’s InhibitorSelect™ Libraries target the same protein (Akt) but show very different chemical structures. Use these molecules to confirm the validity of observed phenotypes.

Structurally diverse. Why is structural diversity so important? Assessing

the effect of multiple, structurally diverse inhibitors

of the same protein target helps rule out potential

non-selective, or “off-target” effects of small

molecules. Non-selective effects typically are seen

when small molecules bind structurally similar

sites on multiple proteins. Inhibitor activity can be

considered “on target” if multiple small molecules

targeting the same protein, but having distinct

chemical structures, all show the same biological

activity.

Further, structurally diverse inhibitors can also help

researchers to gain insight into mechanism of action

of inhibitors. Multiple small molecules with the same

biological activity but possessing distinct chemical

structures targeting different functional domains of

protein, can reveal the biological role of each protein

domain.

Accordingly, our InhibitorSelect™ libraries have been

designed to be structurally diverse, so you can obtain

the maximum interpretable data for each targeted

pathway.

5

InhibitorSelect™ 96-Well Protein Kinase Inhibitor Library I (Catalogue No. 539744)

This panel of compounds consists of 80, well-characterized

protein kinase inhibitors targeting mainly tyrosine, AGC,

and atypical families of kinases, the majority of which are

cell-permeable and ATP-competitive.

InhibitorSelect™ 96-Well Protein Kinase Inhibitor Library II (Catalogue No. 539745)

This panel of compounds consists of 80, well-characterized,

cell permeable, potent and reversible protein kinase

inhibitors targeting mainly CMGC and CaMK families of

kinases; the majority of which are ATP-competitive.

*Read the complete study in EMD Millipore’s Cellutions

2011, Vol. 1, p. 7.

0

2

4

6

8

10

12No GFs

Sub EGF

Sub FGF2

Max GFs

Mouse Neural Stem Cell Viability

SU66

56

GSK

-3 In

hibi

tor X

III

Isog

ranu

latim

ide

IC26

1

IKK-

2 In

hibi

tor I

V

Indi

rubi

n De

rivat

ive

E804

JNK

Inhi

bito

r II

JNK

Inhi

bito

r, N

egat

ive

Cont

rol

JNK

Inhi

bito

r V

JNK

Inhi

bito

r IX

MK2

a In

hibi

tor

JNK

Inhi

bito

r VIII

K-25

2a, N

ocar

diop

sis sp

.

Kenp

aullo

ne

KN-9

3

MEK

Inhi

bito

r I

MEK

Inhi

bito

r II

MEK

1/2

Inhi

bito

r

MN

K1 In

hibi

tor

NK-

kB A

ctiv

atio

n In

hibi

tor

Fold

cha

nge

rela

tive

to D

MSO

Cont

rol G

row

th F

acto

r Bac

kgro

und

Graph (right): InhibitorSelect™ 96-Well Protein Kinase Inhibitor Libraries I & II (160 inhibitors; Cat. Nos. 539744 and 539745) were screened for influence on proliferation and survival of mouse neural stem cells (mNS) in a cell viability assay under 4 conditions: (A) No GFs – No Growth Factors (to identify survival/proliferation factors) (B) Sub EGF – Sub-optimal EGF (to identify inhibitors/potentiators) 20 pg/mL EGF (C) Sub FGF2 – Sub-optimal FGF2 (to identify inhibitors/potentiators) 500 pg/mL FGF2 (D) Max GFs – Maximal EGF + FGF2 (to identify inhibitors/potentiators) 20 ng/mL EGF + 20 ng/mL FGF2 The presence of inhibitor K-252a, Nocardiopsis sp. (Cat. No. 420297) alone in the culture medium resulted in a 10-fold mNS cell viability. Data courtesy of Donna McLaren, Stem Cell Sciences, Cambridge, UK

Screening of 160 kinase inhibitors included in InhibitorSelect™ libraries I and II. Data show Delta Confluence Values, corresponding to the change in relative cell number for twelve mock-treated wells and 160 kinase inhibitors. Three compounds, all affecting Rho kinases, were selected as primary hits for their effect on expansion of Neural Embryonic Stem Cells and are detailed in the top right.

Data courtesy of D Danovi, Wellcome Trust Centre for Stem Cell Research, University of Cambridge

0 1 2Time (days)

Y27632Rho-kinase Inhibitor IVHA 1077 dihydrochloride

Aver

age

delta

con

fluen

ce (n

=3)

3 4100

80

60

40

20

0

-20

1 23

123

InhibitorSelect™ 96-Well Protein Kinase Inhibitor Library III (Catalogue No. 539746)

This panel of compounds consists of 84, well-characterized

protein kinase inhibitors targeting mainly CMGC, CaMK,

AGC, and STE families of kinases, the majority of which are

cell-permeable and ATP-competitive.

InhibitorSelect™ 384-Well Protein Kinase Inhibitor Library I(Catalogue No. 539743)

This panel of compounds consists of 160 well-

characterized, cell-permeable, potent, and reversible

protein kinase inhibitors; the majority of which are ATP-

competitive. This library combines inhibitors from 96-well

Library I and II in one 384-well plate.

0 1 2Time (days)

Y27632Rho-kinase Inhibitor IVHA 1077 dihydrochloride

Aver

age

delta

con

fluen

ce (n

=3)

3 4100

80

60

40

20

0

-20

1 23

123

6

Kinases Targeted By InhibitorSelect™ LibrariesThe table below shows which target kinases are affected by inhibitors present in each of the 4 libraries:

Target Kinases 96-well Library I 96-well Library II 96-well Library III 384-well Library IAdenosine Kinase •

Akt • • •

AMPK • •

ATM • •

ATR • •

Aurora • • • •

Bcr-Abl • •

CaMK • • •

Cdks • • •

cFMS • •

Chk1,2 • • •

CK1,2 • • •

c-Met • •

DAG • • •

DNA-PK • • •

eEF2

EGFR • • •

ERK • •

Flt3 • • •

GSK-3 • •

IGFR • • •

IKK • • •

IP3K •

IRAK • •

JAK • •

JNK • •

Lck • •

MAPK • • •

MEK • • •

MLCK •

MNK1 • •

p70 S6 • •

P90 S6 •

PAK •

PDGFR • •

PDK1 • •

PI 3-K • • •

PIKFyve •

PIM •

PKA •

PKC • • •

PKG •

PKR • •

PLK •

Rho • • •

RIP •

SK • •

Src • • •

Syk • •

TGF-βR • •

Tpl2 •

VEGFR • •

Wee1 •

7

Technology Highlight

The InhibitorSelect™ Protein Kinase Inhibitor Libraries

provide broad coverage of the human kinome as shown

here using the EMD Millipore Data Analysis and Report Tool

(DART™). The depicted human kinome dendrogram of 518

kinases are classified into five broad groups, 90 families,

and 145 subfamilies1. Inhibitor coverage was assigned

based upon published data related to potency (IC50, EC50,

Kd, etc.) for individual kinases harvested from the literature.

Colored dots denote which library contains an inhibitor

with demonstrated potent activity against the designated

kinase and do not necessarily reflect known specificity of

the inhibitor. Coverage of lipid and atypical kinases are

depicted as a separate dendrogram. As shown, Calbiochem®

Protein Kinase Inhibitor Libraries cover all major kinase

families including TK, CMGC, CAMK, AGC, CK1, STE, TKL, as

well as Lipid or Atypical kinase families.

Reference:1. The Protein Kinase Complement of the Human Genome,

G. Manning, D. B. Whyte, R. Martinez, T. Hunter, and S. Sudarsanam, Science 6 December 2002: 298 (5600), 1912-1934.

The DART™ tool – the first software application for in-depth visualization of profiling data.

Visualize the activity and selectivity of your inhibitor

libraries and panels using EMD Millipore’s new Data

Analysis and Report Tool (DART™), which creates an

interactive map of target profiling assay results, enabling

you to make faster, more informed decisions in your drug

development projects.

With the new DART™ Target Visualization Tool, you can: • Securely load previously obtained KinaseProfiler™,

GPCRProfiler™ or AllostericProfiler™ service data

• Map data onto interactive kinome and GPCRome

diagrams

• Visualize % inhibition, % activity and other data

correlated with submitted compounds

• Customize normalization and display options

• Compare activities of up to 4 compounds at once

• Save or print mapped data

Visit www.millipore.com/visualize to try a demo and sign

up to start using DART™.

8

Description Catalogue No.

Phosphatase Inhibitor Cocktail Set II 524625-1SET

Protease Inhibitor Cocktail Set III, EDTA-Free 539134-1SET

Caspase Inhibitor I 627610-1MG

SB 203580 559389-1MG

γ-Secretase Inhibitor XX 565789-500UG

InSolution™ MG-132 474791-1MG

Description Catalogue No.

Anti-Angiopoietin-1 AB10516

Anti-VEGF Receptor-3, extracellular domain, clone 9D9F9 MAB3757

Anti-FGF-2/basic FGF (neutralizing), clone bFM-1 05-117

Flt-1(VEGFR1) active purified kinase 14-562

Anti-Hypoxia Inducible Factor 1a (HIF-1a) 07-628

Anti-PDH-E1a (pSer293) Rabbit pAb AP1062

Description Catalogue No.

Phospho-Histone H2A.X (Ser 139) and p53 QCI / Kit Assay Kit HCS225

Phospho-Histone H3(Ser10) and Cyclin B1 QCI /HCA Assay Kit HCS211

Senescence Detection Kit QIA117 n

BrdU Cell Proliferation Assay, HTS HTS01 n

BrdU Immunohistochemistry System HCS30 n

In Vitro Angiogenesis Assay Kit ECM625

To view our complete Inhibitor Resource page, including product listings, technical tips, and oursubstructure searchable database, please visit www.emdbiosciences.com/Inhibitors.

Best-selling Individual Inhibitors for Analyzing Cell Phenotype

Antibodies

Assays

n Order at www.emdbiosciences.comAll others can be ordered at www.millipore.com

Key Related Products II.

9

Pluripotent embryonic stem cells (ESC) and induced

pluripotent stem cells (iPSC) represent unique and powerful

model systems to study development and differentiation at

the cellular level. Small molecules not only help

identify pathways and mechanisms that control stem

cell fate, they also enable us to manipulate cell fate for

research or therapeutic purposes.

Researchers have successfully used a direct reprogramming

strategy via small molecules to convert mouse fibroblasts

into cardiomyocytes1. It is possible that small molecules

can be used to accelerate manifestation of disease

phenotypes, thereby facilitating revealing of phenotypes

otherwise undetectable in iPSC2. Small molecules have

been used to generate “naïve” human ESCs that can allow

molecular dissection of previously undefined pluripotent

state in humans3. Screening of small molecules can be a

valuable tool in the identification of chemical compounds

that can enrich for cell type of interest in the directed

differentiation of ESCs and iPSC2. Further, small molecule

inhibitors can be used in the induction and long-term self-

renewal of primitive neural precursors from human ESCs4.

References:

1. Efe, J.A. et al. (2011) Conversion of mouse fibroblasts into cardiomyocytes using a direct reprogramming strategy. Nature Cell Biol. published online 30 January 2011; DOI: 10.1038/ncb2164.

2. Wu, S.M. and Hochedlinger, K. (2011) Harnessing the potential of induced pluripotent stem cells for regenerative medicine. Nature cell Biol. 13, 497-505.

3. Hanna, J. et al. (2010) Human embryonic stem cells with biological and epigenetic characteristics similar to those of mouse ESCs. PNAS 107, 9222-9227.

4. Li, W. et al. (2011) Rapid induction and long-term self-renewal of primitive neural precursors from human embryonic stem cells by small molecule inhibitors. PNAS Early Edition, www.pnas.org/cgi/doi/10.1073/pnas.1014041108.

Modulation of cell fate

II.

10

The StemSelect® Small Molecule Regulators 384-Well

Library I consists of 303 pharmacologically active,

structurally diverse small molecules, including extracellular

domain-targeting reagents as well as cell-permeable

compounds that effectively regulate intracellular targets.

The reagents in this library are useful for studying the

survival, migration, proliferation, differentiation, signaling,

and other functions of normal or cancer stem cells as well

as non-stem cells. The library is supplied with a compact

disk containing comprehensive documentation for each

compound.

Convenient Format• Eppendorf® 384-well, Polypropylene, nonpyrogenic,

deep well plate

• Corning® 384-well robolid microplate seal

• Individual silicone seal for each well that exhibits DMSO

resiliency and protects from moisture

• Minimizes cross-contamination

Well-characterized• Cell permeable*

• Potent and selective*

• Reversible*

• Structurally diverse

• Known pharmacological activity

• Stable in DMSO or H2O as supplied

• Less toxic

Comprehensive Documentation• SD Files

• CAS numbers (where available)

• Concentration

• Target

• Categorical index

• PubChem Substance ID (where available)

• Lot specific purity

• Molecular formula

• Molecular weight

• Structure

• Web links to Calbiochem and PubChem for individual

small molecule regulators

*Pertains to a majority of the regulators

Visit www.emdbiosciences.com/StemSelect to learn more

information about StemSelect® Library.

StemSelect® Small Molecule Regulators 384-Well Library I(Catalogue No. 569744)

-50

0

50

100

150

200

1 4 8 12 16 20 24 AD

controlsH

L

P

StemSelect® library molecules affecting cardiomyocyte differentiation

+

-

StemSelect®, a library of highly targeted, well-characterized compounds, provide a high hit rate when screening for modulators of cardiomyocyte differentiation. With three molecules (indicated) showing activity, this library proved more effective than doing a large-scale, untargeted compound screen.Data Courtesy of Dr. Mark Mercola, Sanford|Burnham Institute

11

Summary of Small Molecule Regulators Included in StemSelect™ Library

Small Molecule RegulatorQty

Included

Adenylate Cyclase and Guanylate Cyclase Activators

4

Adenylate Cyclase Inhibitors 2

Adrenergic Receptor Agonists 1

AMPK Activators 3

Angiogenesis Inhibitors 10

Angiogenesis Promoters 1

Anticancer Agents 10

Antioxidants and Free Radical Scavengers 9

Apoptosis Blockers, Other 4

Apoptosis Inducers 3

ATPase Inhibitors 6

Autophagy Inhibitors 1

Calcium Channel Modulators 3

Calpain Inhibitors 2

Cell Adhesion Research Tools, Other 2

Cell Differentiation/Dedifferentiation Inducers

14

Chelating Agents 4

Chemokine Receptor Antagonist 2

c-Myc Inhibitors 1

Coenzymes and Cofactors 2

Cyclooxygenase Inhibitors 12

Cytoskeletal Research Tools 9

DNA and RNA Polymerase Inhibitors 1

DNase and RNase Inhibitors 1

Farenesyl- and Geranylgeranyl-transferase Inhibitors

1

Glucagon Receptor Antagonists 1

Glucocorticoid Receptor Modulators 1

Glutamate Receptor Agonists 1

Glutamate Receptor Antagonists and Uptake Blockers

2

Glycogen Phosphorylase Inhibitor 1

Glycoprotein Processing and Trafficking Inhibitors

1

G-Protein Activators/Modulators 3

G-Protein Antagonists 2

GSK-3 Inhibitors 3

GTPase Inhibitors 2

Heat Shock Protein Inhibitors 2

Histone Acetyltransferase Activators 1

Histone Acetyltransferase Inhibitors 1

Histone Deacetylase Inhibitors 14

HMG-CoA Reductase Inhibitors 3

Hormones, Steroidal 5

Immunomodulators 9

Insulin Secretagogues 3

Small Molecule RegulatorQty

Included

Interleukin Receptor Antagonists 1

Ion Channel Openers 2

Ion Channel Blockers 8

Ionophores 3

IP3 and Ryanodine Channel Modulators 2

Kinase Inhibitors 8

Lipoxygenase Inhibitors 3

Matrix Metalloproteinase Inhibitors 2

Membrane Trafficking/Exocytosis and Endocytosis Products

3

Methyltransferase Inhibitors 5

Mitochondrial Mega Channel Inhibitors and Other Related Products

2

Mitochondrial Metabolism Modulators 1

NF-kB Activation Inhibitors 5

Nuclear Receptor Agonists 3

Nucleic Acids, Nucleotides, Nucleosides, Purines, and Pyrimidines

3

Opioid Receptor Antagonists 1

Other Activators/Agonists 4

Other Inhibitors of Biological Interest 5

p53 Activators 1

p53 Transactivation Inhibitors 2

PARP Inhibitors 5

Phosphatase Inhibitors 10

Phosphodiesterase Inhibitors 6

Phospholipase Activators 1

Phospholipase Inhibitors 4

PPAR Agonists 3

PPAR Antagonists 2

Protease Inhibitors, Other 1

Proteasome-Ubiquitination Inhibitors 5

Protein Kinase C Activators 1

Protein Synthesis Inhibitors 2

Receptor Antagonists, Others 5

Secretase Inhibitors 4

Sonic Hedgehog Signaling Agonists 1

Sonic Hedgehog Signaling Antagonists/Inhibitors

8

Sphingomyelinase Inhibitors 1

STAT Signaling Enhancers 1

STAT Signaling Inhibitors 5

Tautomerase Inhibitors 1

Telomerase Inhibitors 1

TNF- Antagonists 2

Tyrosine Kinase Inhibitors 6

12

Description Catalogue No.

InSolution™ Y-27632 688001-500UG n

Cyclopamine-KAAD 239804 n

TGFβ RI Inhibitor II 616452 n

JAK Inhibitor I 420099 n

Rapamycin 553210 n

Valproic Acid 676380 n

Description Catalogue No.

PDX-1, clone 6F6.1 MAB4425

LEF1, all isoforms, clone 1C3.1D10 MAB3750

BCRP, clone 5D3 MAB4155

BMP7, clone 2A10 MAB4350

Plet1 (Placenta-expressed transcript 1 protein), clone 1D4 MAB4416

SNAI2, clone 2B6 MAB4371

Description Catalogue No.

AXIS™ Axon Investigation System AX45010

QCM™ Chemotaxis Cell Migration Assay, 24-well (8μ), Colorimetric ECM508

QCM™ ECMatrix™ Cell Invasion Assay, 24-well (8μ), Colorimetric ECM550

Calpain Activity Assay Kit, Fluorogenic QIA120 n

Osteogenesis Quantitation Kit ECM815

Human Embryonic Germ Layer Characterization Kit SCR030

Best-selling Individual Inhibitors for Modulating Cell Fate

Antibodies

Assays

iPS Cell Reprogramming Kits and Media

n Order at www.emdbiosciences.comAll others can be ordered at www.millipore.com

Key Related Products

To view our complete Inhibitor Resource page, including product listings, technical tips, and our substructure searchable

database, please visit www.emdbiosciences.com/Inhibitors.

Description Qty/Pk Catalogue No.

Human STEMCCA™ Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

30 μL lentivirus + Polybrene® transfection reagent SCR544

Human STEMCCA™ Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

30 μL lentivirus + Polybrene transfection reagent SCR545

Mouse STEMCCA™ Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

15 μL lentivirus + Polybrene transfection reagent SCR510

Mouse STEMCCA™ Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

45 μL lentivirus + Polybrene transfection reagent SCR530

Mouse STEMCCA™ Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

15 μL lentivirus + Polybrene transfection reagent SCR511

Mouse STEMCCA™ Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

45 μL lentivirus + Polybrene transfection reagent SCR531

ESGRO®-2i Defined Medium containing GSK3β and Mek 1/2 inhibitors to enhance serum-free, feeder-free viability and pluripotency of ES and iPS cells

SF016-100

III.

13

Description Qty/Pk Catalogue No.

Human STEMCCA™ Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

30 μL lentivirus + Polybrene® transfection reagent SCR544

Human STEMCCA™ Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

30 μL lentivirus + Polybrene transfection reagent SCR545

Mouse STEMCCA™ Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

15 μL lentivirus + Polybrene transfection reagent SCR510

Mouse STEMCCA™ Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

45 μL lentivirus + Polybrene transfection reagent SCR530

Mouse STEMCCA™ Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

15 μL lentivirus + Polybrene transfection reagent SCR511

Mouse STEMCCA™ Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

45 μL lentivirus + Polybrene transfection reagent SCR531

ESGRO®-2i Defined Medium containing GSK3β and Mek 1/2 inhibitors to enhance serum-free, feeder-free viability and pluripotency of ES and iPS cells

SF016-100

Signal transduction involves thousands of different

molecules and hundreds of different pathways in the

cell. Small molecules can be used as a valuable tool in

the identification of proteins involved in these different

pathways. Researchers traditionally perform signal

transduction analysis as a linear path, restricting to a

specific pathway in an isolated context within the cell.

More recently, researchers have undertaken a more

system approach involving dynamic interaction networks1.

Such dynamic analysis would integrate high throughput

molecular profiling, database and literature mining,

mechanistic modeling, and cell culture experiments.

Reference:

1. Kirouac, D.C. et al (2010) Dynamic interaction networks in a hierarchically organized tissue. Mol. Syst. Biol. 6, 1-16.

InhibitorSelect™ Signaling Pathway PanelsRecognizing both the tremendous opportunities and the

challenges in analyzing signal transduction, we have

introduced several Calbiochem® Signaling Pathway

Panels that contain a collection of carefully selected, cell

permeable, and potent small molecules in convenient

formats. These panels allow the researcher to zero in on

their pathway of interest and gain useful information by

manipulating specific proteins or families.

Analysis of specific signaling pathways

III.

In the following pages, we have highlighted ten Signaling pathway panels:

Visit www.emdbiosciences.com/inhibitorselect to learn more about signaling pathway panels.

1. PI 3-K/Akt/mTOR

2. EGFR

3. JAK/STAT

4. NF-kB

5. MAP Kinase

6. VEGF

7. Wnt

8. IGF

9. FGF

10. Hedgehog

14

1. InhibitorSelect™ PI 3-K/Akt/mTOR Signaling Pathway Inhibitor Panel (Catalogue No. 124031)

Components Target Amount Catalogue No.

Akt Inhibitor IV Akt 1 mg 124011Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 Akt1, Akt2 1 mg 124018LY 294002 PI 3-K 5 mg 440202LY 303511 Negative control for LY 294002 1 mg 440203PDK1/Akt/Flt Dual Pathway Inhibitor Akt, Flt, PDK1 5 mg 521275PI-103 DNA-PK, PI 3-K, mTOR 1 mg 528100PI 3-Kγ Inhibitor PI 3-Kγ 5 mg 528106PI 3-Kγ Inhibitor II PI 3-Kγ 5 mg 528108PI 3-K Inhibitor IV PI 3-K + β 5 mg 528111PI 3-K Inhibitor VIII DNA-Pk, PI 3-K 5 mg 528116Rapamycin mTOR, p70 S6 100 μg 553210Ro-31-8220 PKC 500 μg 557520Wortmannin PI 3-K Irreversible 1 mg 681675DMSO - 15 mL KP31817

IGF-1

IGF-1R

PKCα

Growth Factors

RTKs

JAK1

PI 3-K

• LY 294002 • PI-103 • PI 3-Kγ Inhibitor • PI 3-Kγ Inhibitor II • PI 3-Kα Inhibitor IV • PI 3-Kα Inhibitor VIII • Wortmannin

• PDK1-Akt-Flt Dual Pathway Inhibitor • Akt Inhibitor IV, Akt Inhibitor VIII

• PDK1-Akt-Flt Dual Pathway Inhibitor

• PI-103, PI 3-Kα Inhibitor VIII

• Ro-31-8220

JAK1GAB1 BCAP

MDM2

MDM2Nucleusp53

HSP90CDC37

Cytokines AG

Cytokine Receptor BCRBCR

4EBP1

elF4E

PDK-1

Raf1

• PI-103 • Rapamycin

mTor

p70s6K

PP2A

DNA-PK

SYK

Caspase 9

Caspase Cascade

ERK Pathway

Translation

Protein Synthesis

p53 Degradation

GlycogenSynthesis

GSK-3

Akt

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

Activation of the PI 3-kinase/Akt/mTOR pathway stimulates cell proliferation and the translation process in response

to nutrients and growth factors. Dysregulation of this pathway can lead to a variety of human tumors. Over 30% of

solid tumors are reported to contain mutations in the catalytic unit of their PI 3-K, resulting in increased enzymatic

activity, cell proliferation, cell invasion, and metastasis. The InhibitorSelect™ PI 3-K/Akt/mTOR Signaling Pathway Inhibitor Panel enables multiparameter analysis, assessment of signal amplification/feedback, and comparison of

biological effects of perturbing different parts of the pathway.

15

2. InhibitorSelect™ EGFR Signaling Pathway Inhibitor Panel (Catalogue No. 324839)

Components Target Amount Catalogue No.

Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 Akt1, Akt2 1 mg 124018PD 153035 EGFR 1 mg 234490EGFR Inhibitor EGFR, EGFR Mutants 1 mg 324674Et-18-OCH3 PI-PLC 5 mg 341207JNK Inhibitor II JNK 5 mg 420119LY 294002 PI 3-K 5 mg 440202PD 98059 MEK 5 mg 513000PD 168393 EGFR Irreversible 1 mg 513033PP2 Src Family 1 mg 529573Rapamycin mTOR, p70 S6 100 μg 553210SB 203580 p38 MAPK 1 mg 559389AG 490 JAK2, EGFR 5 mg 658401ZM 336372 c-Raf 1 mg 692000DMSO - 15 mL KP31817

EGF

EGFRErbB2

Nucleus

SHCGRB2

GAB1 DOK2

Rac1

EPS8Muc1

Ctnnβ

GβL

p70S6K

Raptor

SOSPLCγ FAK1

PKC

Ras

Raf

MEK1/2

ERK1/2

p120GAP

c-Src

PAK1 JAK1/2c-Src

MEKK1

MKK4/7

JNKsCSK

IKK

PDK-1

Akt

mTor

IP3DAG PIP2

PIP3

• PD 153035 • EGFR Inhibitor

• PD 168393 • AG 490

• Akt Inhibitor VIII• Isozyme-Selective

Akt1-1/2

• LY 294002

• ZM 336372

• Rac1 Inhibitor

• PP2

• AG 490

• Rapamycin

• Et-18-OH3

• PD 98059 • PD 98059

• JNK Inhibitor II

mTORC1

Translation

AKT Signaling

Cell Survival

MAPKSignaling

Gene Expression

FAK Pathway

Cell Motility

CytoskeletonRegulation

IP3Signaling

STAT1PI 3-K

Vav

STAT1STAT3

STAT1STAT3

STAT3

JAK1/2

c-Myc

STAT1STAT1

NF-κB

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

The epidermal growth factor receptor (EGFR) is a transmembrane receptor tyrosine kinase involved in cell

proliferation, growth, migration, invasion, and survival. EGFR is overexpressed in a vast majority of human epithelial

tumors and constitutes an attractive target for the development of novel anticancer agents. EGFR is a receptor that

integrates so many diverse signals and has so many different outputs, that only by using a panel of small molecules,

like the InhibitorSelect™ EGFR Signaling Pathway Inhibitor Panel, can you truly understand the phenotypic

impact of perturbing EGF signaling in a given cellular environment.

16

Components Target Amount Catalogue No.

PD 153035 EGFR 1 mg 234490Indirubin Derivative E804 Src, Cdk1/cyclin E, Cdk2/cyclin A, Cdk1/cyclin B 1 mg 402081JAK Inhibitor I JAK1, JAK2, JAK3, Tyk2 500 mg 420099JAK2 Inhibitor II JAK2 25 mg 420132LY 294002 PI 3-K 5 mg 440202PP2 Src 1 mg 529573SHP1/2 PTPase Inhibitor, NSC-87877 SHP1, SHP2 50 mg 565851STAT3 Inhibitor Peptide, Cell Permeable STAT3 1 mg 573096STAT3 Inhibitor III, WP1066 STAT3 10 mg 573097STAT3 Inhibitor V, Stattic STAT3 25 mg 573099STAT5 Inhibitor STAT5 10 mg 573108AG 490 JAK2 5 mg 658401U0126 MEK1, MEK2 1 mg 662005DMSO - 15 ml KP31817

Nucleus

Growth Factors

RTK

Growth Hormones

Growth Hormone Receptors

JAKsJAKs

Cytokines GPCR Ligands

Cytokine Receptor GPCR

• AG 490 • JAK2 Inhibitor II • JAK Inhibitor I

• PP2, Indirubin Derivative

• STAT5 Inhibitor

• STAT5 Inhibitor

• STAT3 Inhibitor V, Stattic • STAT3 Inhibitor Peptide, Cell Permeable • STAT3 Inhibitor III, WP1066

• STAT3 Inhibitor V, Stattic • P3-AHNP Peptide Carrier• STAT3 Inhibitor III, WP1066

• STAT3 Inhibitor V, Stattic • P3-AHNP Peptide Carrier• STAT3 Inhibitor III, WP1066

• SHP1/2 PTPase Inhibitor, NSC-87877

• AG 490• JAK2 Inhibitor II,

• JAK Inhibitor I

• AG 490 • JAK2 Inhibitor II • JAK Inhibitor I

• c-Myc Inhibitor

• PD 153035

• LY 294002

• U0126

Akt Pathway

Gene Expression

STAT3STAT5

JAK2

Src

JAKs

Akt

Raf

MEK

ERKs

SHP2JAKs JAKs

Cofactorsc-MycCTFs

STATsSTATs

STAT3STAT3

STATs

STATs STATs

STATs

STATsSTATs

STATsSTATs

STATsSTATs

STAT5STAT5

STATsSTATs

JAK2

Sumo SumoSumo

RANKPNA1

STAM

SH2B

PI 3-K

GRB2SOS Ras

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

3. InhibitorSelect™ JAK/STAT Signaling Pathway Inhibitor Panel (Catalogue No. 420138)

The Janus kinase (JAK) signal transducers and activators of transcription (STAT) play an important role in cell

proliferation, cell differentiation, cell migration, and cell death. The JAK/STAT signaling pathway is the principal

signaling mechanism for a variety of cytokines and growth factors. Constitutive activation or dysregulation of JAK/

STAT signaling can result in inflammatory disease, erythrocytosis, gigantism, and leukemia. Because many types of

stimuli can activate JAK/STAT signaling, probing cells with several different combinations of JAK and STAT inhibitors,

such as those included in the InhibitorSelect™ JAK/STAT Signaling Pathway Inhibitor Panel, is the key to

identifying the precise pathway used to transduce the signal of interest.

17

Components Target Amount Catalogue No.

BAY 11-7082 IkB 10 mg 196870IKK Inhibitor VII IKK 1 mg 401486IRAK 1/4 Inhibitor IRAK1, IRAK4 5 mg 407601Histone Acetyltransferase Inhibitor II p300/CBP Histone Acetyltransferase 10 mg 382110Trichostatin A, Streptomyces sp. Histone Deacetylase 1 mg 647925MG-132 Proteasome 1 mg 474790NEMO Binding Domain Peptide, Cell-Permeable NEMO/IKK (IkB)-kinase complex 500 μg 480025NF-kB SN50 NF-kB nuclear translocation 500 μg 481480PD 98059 MEK 5 mg 513000PDK1/Akt-Flt Dual Pathway Inhibitor Akt, PDK1, Flt 5 mg 521275TIRAP Inhibitor Peptide NF-kB, PKR, JNK 1 mg 613570Tpl2 Kinase Inhibitor Tp12 Kinase 1 mg 616373(5Z)-7-Oxozeaenol, Curvularia sp. TAK1, MEK1, MEKK1, ASK1 1 mg 499610JNK Inhibitor II JNK1 5 mg 420119DMSO - 15 mL KP31817

Nucleus

Bacterial Antigens

Growth Factors

Growth FactorReceptors

TNFα

Proteosome

TNFR1 IL-1R

IL-1

• PDK/Akt/Flt Dual

PathwayInhibitor

• Interluekin-1 Receptor-AssociatedKinase-1/4 Inhibitor

• PD 98059

• (5Z)-7-Oxozeaenol, Curvularia sp.

• IKK Inhibitor VII

• IKK Inhibitor VII

• NEMO-Binding Domain BindingPeptide, Cell Permeable • BAY 11-7082

• Tpl2 Kinase Inhibitor

• Histone Acetyltransferase Inhibitor II• Trichostatin A,Streptomyces sp.

• JNK Inhibitor II

• MG-132

• BAY 11-7082

p52RelB

NF-κB

p52p54

NF-κBNF-κB

p50c-Rel

NF-κB

p50c-Rel

NF-κB

IκBs

HDAC3

p50

p50

p50Bcl3

p50

p105p105

ABIN-2Tpl2

LPS

TRADDTRAF2

p300

CBP

TRAF6

TRAF6

MALPPGN

MyD88TOLLIPMyD88

PI 3-K

PDK-1

ASK1

MKK4,7

JNKsAkt/PKB

IKK-γ

MEKKs

Tpl2

MEKK

ERK

PKR PKR

IRAKIRAK

IRAK

TAK1

IKK-α IKK-β

JNK PathwayERK

Signaling

Tpl2Degradation

Virus

IκB Degradation

Nuclear Import

SCF/β-TrCP

IκBs

RelBp52NF-κB

IκBs

IκBs

p50c-Rel

NF-κB

IκBs p52p65

NF-κB

κ-BRas

IκBs p52p65

NF-κB

IκBs p52p65

NF-κB

PACT

Nuclear Export

AcetylationDeacetylation

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

4. InhibitorSelect™ NF-kB Signaling Pathway Inhibitor Panel (Catalogue No. 481487)

The eukaryotic nuclear factor kB (NF-kB) plays an important role in inflammation, autoimmune response, cell

proliferation, and apoptosis by regulating the expression of genes involved in these processes. The Rel/NF-kB signal

transduction pathway is mis-regulated in a variety of human cancers, especially those of lymphoid cell origin.

Designing antitumor agents to block NF-kB activity or to increase their sensitivity to conventional chemotherapy

may have great therapeutic value. Basing such target validation studies on a set of structurally diverse molecules

such as in the InhibitorSelect™ NF-kB Signaling Pathway Inhibitor Panel can provide a powerful starting point for

drug discovery.

18

Components Target Amount Catalogue No.

ERK Inhibitor II, FR180204 ERK1, ERK2 1 mg 328007JNK Inhibitor II JNK 5 mg 420119JNK Inhibitor IX JNK2, JNK3 5 mg 420136MEK1/2 Inhibitor MEK1/2 1 mg 444939MNK1 Inhibitor MNK1 5 mg 454861MK2a Inhibitor MK2a 5 mg 475863p38 MAP Kinase Inhibitor V p38, CK1 1 mg 506156PD 98059 MEK 5 mg 513000Raf Kinase Inhibitor IV B-Raf 1 mg 553014SB 203580 p38 MAPK 1 mg 559389Tpl2 Kinase Inhibitor Tpl2 Kinase 1 mg 616373ZM 336372 c-Raf 1 mg 692000DMSO - 15 mL KP31817

Nucleus

Growth Factors

RTK

Hormone Cytokines HyperosmoticShock

GPCR

• MEK1/2 Inhibitor, PD 98059

• ERK Inhibitor II,FR180204

• JNK Inhibitor II• JNK Inhibitor IX

• PD 98059

• p38 MAP KinaseInhibitor V,SB 203580

• Tpl2 Kinase Inhibitor

• Raf Kinase Inhibitor IV

• MNK1 Inhibitor

• MK2a Inhibitor

• ZM 336372• Raf Kinase Inhibitor IV

• PD 98059

Cell Proliferation, Cell Survival,Tumorigenesis, Differentiation,Development

Gene Expression

Gene ExpressionApoptosis, Inflammation,

Tumorigenesis

Cell Motility, Inflammation,

Apoptosis, Osmoregulation

Gα SHC SOS

GRB2

Gγ Gα

Gβ

PLC

Rac1

RAS

Hormones

N

C

GPCR

N

C

TNF

TNFRTLR4

LPS

IL-1R

IL1

OSM

PLC

DAG DAGcAMP

PKC

c-Raf

Tpl2 MAP3Ks

MEK4/7

JNK1/2/3JNK1/2/3 p38

p38

RIP

RAFMEK1/2

MEK3/6MAP3KsPKCPKA

AC

ERK1/2ERK1/2

MSK RSK

CDC42

TRADD

TRAF2

IRAKTRAF6

c-Junc-Fos

STATsElk1

c-Myc

CREB MNK CREB

p53MAPKAPK2

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

5. InhibitorSelect™ MAP Kinase Signaling Pathway Inhibitor Panel (Catalogue No. 444189)

The mitogen-activated protein (MAP) kinases are a group of evolutionarily conserved protein serine/threonine kinases

that are activated in response to a variety of extracellular stimuli. Together with several other signaling pathways

they can differentially alter the phosphorylation status of various transcription factors. A controlled regulation

of these cascades is involved in cell proliferation and differentiation, whereas an unregulated activation of these

MAP kinases can result in oncogenesis. The InhibitorSelect™ MAP Kinase Signaling Pathway Inhibitor Panel can

help deconvolute the roles of particular pathway proteins more rapidly than using individual MAP kinase pathway

inhibitors one at a time.

19

Components Target Amount Catalogue No.

Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 Akt1/2 1 mg 124018Calcineurin Autoinhibitory Peptide, Cell-permeable Cal AI Pep 1 mg 207001cPLA2a Inhibitor cPLA2 500 μg 525143ET-18-OCH3 PLC 5 mg 341207Gö 6983 PKC isozymes 500 μg 365251NG-Nitro-L-arginine Methyl Ester, Hydrochloride eNOS 100 mg 483125p21-Activated Kinase Inhibitor III, IPA-3 PAK 5 mg 506106PI-103 PI 3K, mTOR 1 mg 528100PP2 Src family, PAK 1 mg 529573SB 203580 p38 MAPK 1 mg 559389U0126 MEK1/2 1 mg 662005VEGFR Tyrosine Kinase Inhibitor V VEGFR1/2/3 5 mg 676501VEGFR2 Kinase Inhibitor III VEGFR2 non selective 1 mg 676487VEGFR2 Kinase Inhibitor VI, Ki8751 VEGFR2 5 mg 676484ZM 336372 c-Raf 1 mg 692000DMSO - 15 mL KP31817

VEGFA, VEGFB

VEGFR1

VEGFA, VEGFC, VEGFD

VEGFR2

VEGFC, VEGFD

VEGFR3• VEGFR Tyrosine Kinase

Inhibitor V

• Akt Inhibitor VIII• Isozyme-Selective, Akti-1/2

• PP2 • p21-Activated

KinaseInhibitor III,

IPA-3

• NG-Nitro-L-arginine Methyl Ester, Hydrochloride

• VEGFR2 Kinase Inhibitor III• VEGFR Tyrosine Kinase Inhibitor V

• VEGFR2 Kinase Inhibitor VI, Ki8751

• PI-103

• PI-103

• SB 203580

• cPLA2a Inhibitor

• Go 6983

• U0126

• ZM 336372

• ET-18-OCH3

• CalcineurinAutoinhibitory Peptide,

Cell-Permeable

• VEGFR TyrosineKinase Inhibitor V

Survival

HSP27

Cell Survival

NFAT Signaling

ProstaglandinProduction

Gene Expression &Cell Proliferation

Focal Adhesion

Vascular Cell PermeabilityAngiogenesis

PIP3 PIP2 PIP2

DAG Ca2+

Nitric OxideProduction

Src

PLCγ

MKK3/6 PKC Calcineurin

p38Raf1

MEK1/2 ERK1/2

cPLA2

Akt/PKB

mTOR

Caspase 9

eNOSNFAT

PI 3-KVRAPSck

Actin Reorganization

Ras

HSP90

PAKNCK SOSSHC

GRB2

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

6. InhibitorSelect™ VEGF Signaling Pathway Inhibitor Panel (Catalogue No. 676502)Vascular endothelial growth factor (VEGF) family of proteins have been implicated in the development, maintenance,

and remodeling of vasculature and other physiological processes such as endothelial cell growth, monocyte migration,

tumorigenesis, lymphangiogenesis, and the paracrine release of growth factors from endothelial cells. Mis-regulation

within the VEGF family has been implicated in a variety of other human diseases, including metastasis, hyperthyroidism,

atherosclerosis, and inflammatory diseases, such as rheumatoid arthritis and psoriasis. The InhibitorSelect™ VEGF Signaling Pathway Inhibitor Panel enables simultaneous, dynamic study of multiple signaling pathways activated

by VEGF.

20

Components Target Amount Catalogue No.

Casein Kinase I Inhibitor, D4476 Casein Kinase I 1 mg 218696Casein Kinase II Inhibitor III, TBCA Casein Kinase II 5 mg 218710β-Catenin/Tcf Inhibitor, FH535 β-Catenin, Tcf 10 mg 219330GSK-3 Inhibitor IX GSK-3 1 mg 361550PKG I Inhibitor, Cell-Permeable PKG 1 mg 370655H-89, Dihydrochloride PKA, Rho Kinase 1 mg 371963JNK Inhibitor II JNK 100 μg 420119K-252a, Nocardiopsis sp. Multiple 5 mg 420298KN-93 CaMK 1 mg 422708LY 294002 PI 3-Kinase 5 mg 440202(5Z)-7-Oxozeaenol, Curvularia sp. TAK 1 1 mg 499610PP2 Src Family 1 mg 529573Rapamycin mTOR 100 μg 553210SB 202190 p38 1 mg 559388U0126 MEK 1 mg 662005DMSO - 15 mL KP31817

Nucleus

Off-State On-State

Frizzled Frizzled Frizzled Frizzled

• Casein Kinase II Inhibitor III, TBCA

• LY 294002 • PP2

• U0126

SB 202190

• Rapamycin

• PKG Ia Inhibitor,Cell-Permeable

• K-252a, Nocardiopsis sp.

• KN-93; K-252a, Nocardiopsis sp.

• JNKInhibitor

• H-89, Dihydrochloride

• (5Z)-7-Oxozeaenol,Culvaria sp.

• GSK-3 Inhibitor IX

• Casein Kinase I Inhibitor, D4476

• K-252a, Nocardiopsis sp.• H-89, Dihydrochloride

• β-Catenin/Tcf Inhibitor, FH535

NFAT

CaderinCer

Caderin

APCAXIN

DSH TAB DSH

Rac1

α-Cateninβ-Catenin

β-Catenin

β-Catenin

GSK-3b

CK1 CKII

GSK-3β

TAK1 PI3-K

Akt

Src

ERKS MAP3Ks

CaMKII

PLCMEKKS

p38

CalnPKC

PDE

PKG

MAPKK4/7Rho Kinase

JNK

mTORNLKPKA

Proteosome

ERK Pathway

CellPolarity

Cell Survival

NFATPathway

PKCPathway

MAPKCascade

Cell Survival/Protein Synthesis

Gene Expression Gene ExpressionCell Fate Proliferation,Differentiation,

Adhesion, and Survival

APCAXIN

β-Catenin

RhoA

β-Catenin

β-Catenin

β-Catenin Degradation

Ca2+

TCF/LEFCREB

ATF2

c-Myc

WNTWNT5A

WNT1/11 WNT5

G Proteins

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

7. InhibitorSelect™ Wnt Signaling Pathway Inhibitor Panel (Catalogue No. 681666) The Wnt signaling pathway is an evolutionarily-conserved pathway involved in fate specification, development,

cell proliferation, cell migration, and polarity, and migration of cells. Wnt genes encode a large family of secreted,

cysteine-rich proteins that are also important in development and in maintenance of adult tissues. Abnormalities

in Wnt signaling are reported to promote both human degenerative diseases and cancer. Like many developmental

pathways, Wnt signaling has a great deal of built-in redundancy; therefore, studying it requires a panel of

small molecules, such as the InhibitorSelect™ Wnt Signaling Pathway Inhibitor Panel for accurate, complete

perturbation.

21

Components Target Amount Catalogue No.

AG 1024 IGF-1R 1 mg 121767Akt Inhibitor IV PDK-1 1 mg 124011Akt Inhibitor VIII, Isozyme selective, Akti-1/2 Akt1/2 1 mg 124018AMPK Inhibitor, Compound C AMPK 1mg 171260bpV(phen) PTEN 10 mg 203695ERK Inhibitor II, FR180204 ERK1/2 1 mg 328007GSK-3β Inhibitor VIII GSK 3β 5 mg 361549IGF-1R Inhibitor, PPP IGF-1R 1 mg 407247NBI-31772 IGFBPs 5 mg 479830PD 98059 MEK 5 mg 513000PI-103 PI 3-K, mTORC1/2 1 mg 528100PKCβ Inhibitor PKCβ 500 μg 539654Rapamycin mTOR 100 μg 553210RSK Inhibitor, SL0101 P90RSK 1 mg 559285ZM 336372 c-Raf 1 mg 692000DMSO - 15 mL KP31817

Nucleus

GLUT4

• Rapamycin• PI-103

• GSK-3β Inhibitor VIII

• Rapamycin • PI-103

• PI-103

• PKCβ Inhibitor

• AMPK Inhibitor, Compound C • RSK Inhibitor

SL0101

• Akt Inhibitor IV

• ERK Inhibitor II, FR180204

• Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2

NF-κB

• ZM 336372

• PD 98059

• AG 1024 • IGF-1R Inhibitor, PPP• NBI-31772• bpV(phen)

GβLRaptor

PI 3-K

PTEN

PDK-1PKCλ

GSK3β

GlycogenSynthase

PKCζ

mTORGβL

RictormTOR

AMPK

Raf

MEK1/2 PKCβ Calm

CalmKs

Calcineurin

ERK1/2AKT

IKKs

p90RSK

Caspase 9

Crk

Glycogen Sythesis

Cell Survival

Proliferation &Differentitation

GlucoseUptake

Gene Expression

IκB Degradation

ProteinSythesis

GLUT4

IGF1, IGF2

IGF1R

PIP2PIP3

GLUT4Vesicle

elF4E

elF2B

NFAT

NFAT

FKHRL1

IκB

IRS SHCGRB2

SOS

IGFBPs

C3G

Ras

Ca2+

Ca2+

mTORC1

mTORC2 NF-κB

NFAT

CREBElk1

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

8. InhibitorSelect™ IGF Signaling Pathway Inhibitor Panel (Catalogue No. 407249)

The IGF receptors serve as primary mediators of endocrine as well as autocrine and paracrine actions that promote

cell proliferation, survival and differentiation in pre- and post-natal development. These receptors contribute

to mammary epithelial stem cell maintenance and renewal, progenitor cell expansion, and motility among

neuroblastoma cancer cell lines. Misregulation of receptor and ligand expression and their cross interaction has been

associated with several types of cancer. Using a panel of inhibitors, such as the InhibitorSelect™ IGF Signaling Pathway Inhibitor Panel can accelerate studies of IGF signaling by decoupling, for example, receptor binding events

from downstream signaling.

22

Components Target Amount Catalogue No.

Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 Akt 1/2 1 mg 124018EGF/FGF/PDGF Receptor Tyrosine Kinase Inhibitor FGFR-1, PDGFR-β, c-Src, and EGFR 2 mg 324841ET-18-OCH3 PLC 5 mg 341207FGF/VEGF Receptor Tyrosine Kinase Inhibitor, PD173074 FGFR > VEGFR 5 mg 341607FGF Receptor Tyrosine Kinase Inhibitor FGFR 5 mg 341608Gö 6983 PKC 500 μg 365251JNK Inhibitor VIII JNK 1, 2, 3 5 mg 420135LY 294002 PI 3-K 5 mg 440202PD 98059 MEK/ERK 5 mg 513000PP2 PAK 1 mg 529573Rac 1 Inhibitor Rac1 5 mg 553502STAT3 Inhibitor III, WP1066 STAT3 10 mg 573097U0126 MEK1/2 1 mg 662005ZM 336372 c-Raf 1 mg 692000DMSO - 15 mL KP 31817

FGF

FGFR

• Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2

• LY 294002

• PP2 PD-089828

• Et-18-OCH3

• PD98059

• PD98059

• JNK Inhibitor VIII

• Go 6983

• ZM 336372

• U0126

• PD-089828• PD 166866• PD173074

• PP2• Rac1 Inhibitor

• WP1066

Cell Survival

Actin Crosslinking

Cell Migration

Neuritogenesis

Invasion,Migration

ProliferationNeurite Outgrowth

GRB2SOS

Rac1

F-Actin

PIXAPPI

PAF

PIP2

DAG

PDK-1

Src Ras

Raf1 PAK

PLA2

Src

ERK1/2MEKsMEKs

MKK3/6

p38

SEK

JNK

PKC

Akt

STAT3

PI 3-K

FRS2 PLCγ

ATF2Elk1

NucleusPhosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

9. InhibitorSelect™ FGF Signaling Pathway Inhibitor Panel (Catalogue No. 341612)

Fibroblast Growth Factors (FGF) are heparin-binding proteins, which interact with cell-surface associated heparin

sulfate proteoglycans to produce a wide array of cellular and physiological effects, such as angiogenesis, wound

healing, and embryonic and neural development. Mis-regulation of FGF signaling has been closely associated with

various human diseases, such as Michel aplasia, neoplasia, Parkinson disease, Pheiffer syndrome and hereditary

spinocerebellar ataxias. The InhibitorSelect™ FGF Signaling Pathway Inhibitor Panel provides a means to

simultaneously assess the effects of FGF signaling on diverse biological processes.

23

Components Target Amount Catalogue No.

Smoothened Agonist, SAG Smo agonist; Hh pathway activator 1 mg 566660Cyclopamine-KAAD Smo antagonist 100 μg 239804Hh Signaling Antagonist XIV, SANT-2 Smo antagonist, targets different site than Cyclopamine 10 mg 373273Hh/Gli Antagonist, GANT61 Gli antagonist 5 mg 373401GSK-3 Inhibitor IX, BIO Reduces Gli1-dependent transcriptional activity 1 mg 361550H-89, Dihydrochloride PKA Inhibitor; Hh agonist 1 mg 371963IC261 CK1 Inhibitor (CK1δ and CK1ε; IC50 = 0.7-1.3 μM and

= 0.6-1.4 μM, respectively)5 mg 400090

GRK2 Inhibitor Reduces phosphorylation of human Smo 5 mg 182200MG-132 Proteasomal Inhibitor 5 mg 474790Hh Signaling Antagonist VII, JK184 Targets Adh7, inhibits Gli-transcription activity, and down-

regulates the expressions of Gli1 and Ptc15 mg 373385

Purmorphamine Smo agonist, targets Cylopamine binding site and upregulates Gli1 and Ptc1

5 mg 540220

Fluvastatin, Sodium Salt Cholesterol biosynthesis inhibitor 25 mg 344095Hh Signaling Antagonist XII, HPI-1 Remains effective against Sufu-/- fibroblasts overexpressing

Gli1 or Gli210 mg 373275

Hh Signaling Antagonist XIII, HPI-3 Ineffective against Sufu-/- fibroblasts over-expressing Gli1 or Gli2

10 mg 373276

DMSO - 15 mL KP31817

Nucleus

Nucleus

Hedgehog Secreting Cell

Hedgehog Receiving Cell

• GRK2 Inhibitor

• H-89, Dihydrochloride

• IC261 • H-89, Dihydrochloride

• Fluvastatin, Sodium Salt

• Smoothened Agonist, SAG • Purmorphamine

• Cyclop-KAAD• Hh Signaling Antagonist XIV, SANT-2

• IC261

• MG-132

• Hh/Gli Antagonist• GAN61

• Hh Signaling Antagonist VII, K184• Hh Signaling Antagonist XIII, HPI-3• GSK-3 Inhibitor IX, BIO• Hh Signaling Antagonist XII, HPI-1

Hh

Hh

Hedgehog

Proteosome

Secretion

Hh Precursor

-Hh

Target Genes(WNT, Ptc, BMP)Target Genes

+Hh

Gli2Degradation

N C

Cholesterol

Palmitate

N C

Ptc

N C

Ptc

N

C

Smo

SUFUGli 2/3

Gli SUFU

Microtubule

Gli

Gli 2/3GSK3β

PKA

CK1

AC

PKA

STK36

GRK CK1 GSK3β

G Proteins

GliCBPCoRCoR

GliR

GliR

Phosphorylation

Ubiquitin

GTP

Calbiochem® Inhibitors

10. Hedgehog Signaling Pathway Modulators Panel (Catalogue No. 373386)Hedgehog signaling, a key intercellular signaling pathway in differentiation and organogenesis, is conserved from

flies to human. Vertebrates express three different hedgehog proteins, of which Sonic hedgehog (Shh) is the

best characterized. Shh has been implicated in several embryonic developmental processes. It displays inductive,

proliferative, neurotrophic, and neuroprotective properties. Mutations in the Shh pathway can lead to congenital

defects and diseases, including cancer. Because Hh signaling involves both intercellular and intracellular signaling,

EMD Millipore’s Hh Signaling Pathway Modulators Panel provides a convenient way to simultaneously interrogate

signal secreting and signal receiving cells in one experiment.

24

Turn hits into candidates with Lead Discovery Profiling ServicesIn addition to revealing cellular phenotypes and signaling

pathways in research studies, many small molecule

inhibitors and activators have been successfully been

developed as therapeutic drugs. EMD Millipore’s Lead

Discovery Services are well suited to extensive profiling

of these drugs against a broad range of targets to reveal

interactions that may have additional therapeutic value

or present adverse reactions. Our lead discovery scientists,

backed by cutting-edge profiling tools and decades of

expertise, provide a perfect complement to your in-house

studies, turning hits into candidates.

GPCRProfiler ServiceGPCRProfiler™ is the first complete cell-based functional

platform that uses a common validated readout for over

155 GPCRs.

Quickly implement screening in a new disease area or

target receptor, or increase your current screening capacity

with GPCRProfiler™ services. Our unbiased profiling

screens can determine if drugs are full or partial agonists

Visual display using EMD Millipore’s DART™ (Data Analysis and Reporting Tool) of the inhibitory activity of the anti-cancer agent imatinib (Gleevec®, Novartis) which was functionally profiled against a large portion of the kinome using KinaseProfiler™ service.

or antagonists. We also perform dose-response assays

to determine EC50 values for agonists and IC50 values

for antagonists. You will typically receive results within

1-3 weeks of compound submission, in a thorough and

intuitive report. GPCRProfiler™ gives you the flexibility to

choose from 1 to ≥155 different receptors to screen. Or,

choose between 15 different service panels, including more

than 60 distinct ligand families, 2 safety panels and 11

disease focused panels.

Kinase & Phosphatase Profiling ServicesIn 2000, the KinaseProfiler™ service developed by

Upstate, now a part of EMD Millipore, brought selectivity

profiling to kinase drug discovery researchers. Today, the

KinaseProfiler™ panel includes almost 300 protein and

lipid kinases, 21 phosphatases and a complementary suite

of secondary assays, forming the most diverse, disease

relevant panel available commercially. As the partner of

choice for kinase screening, we provide validated data

using the robust and reliable radiometric kinase assay

trusted by the world’s leading pharmaceutical companies.

SignalProfiler™ ServicesProviding fresh insights to the cellular activity of

compounds, SignalProfiler service employs MILLIPLEX®

MAPmate™ cell signaling kits to profile samples against a

panel of 90+ modified and total proteins. SignalProfiler™

can be used to measure the effectiveness of kinase

inhibitors and GPCR ligands by measuring changes in

signaling cascades in relevant cell backgrounds. In addition,

early signaling events leading to cellular toxicity can be

studied to help assess potential compound liabilities in

early stages of drug development.

UbiquitinProfiler™ ServiceUbiquitinProfiler™ service is the first service to

provide compound screening and profiling against E3

ligase cascades. Each ubiquitination cascade on the

UbiquitinProfiler™ panel consists of the three enzyme

components (E1 activating enzyme, E2 conjugating enzyme

and E3 ligase), plus a biologically-relevant substrate.

The incorporation of ubiquitin into the substrate is

quantitatively detected by electrochemiluminescence,

which allows for the identification of compound inhibitors

and activators.

Visit www.millipore.com/leaddiscovery to choose and

order profiling services.

Technology Highlight

25

Description Catalogue No.

Chk2 Inhibitor II 220486

SB 218078 559402

Staurosporine 569397

Cdk1 Inhibitor IV, RO-3306 217699

TAS-301 608050

VEGF Inhibitor, CBO-P11 676496

Description Catalogue No.

Anti-Fas (human, activating), clone CH11 05-201

Anti-Cdk5, clone DC17 05-364

Anti-Bcl2, clone 100 05-729

Anti-phospho-erbB-2/HER-2 (Tyr1248) 06-229

Cell Cycle-G2/M Phase Pathway Explorer Antibody Minipack 15-120

Fluorescent Conjugated Antibodies for Cell Surface CD Markers FCMAB211F, FCMAB188F and others

Description Catalogue No.

PI3-Kinase HTRF Screening Assay 33-016

MILLIPLEX® map MAPK/SAPK Cell Signaling 10-Plex 48-660

Akt (Ser473) Dual Detect CELISA Assay Kit (Fluorogenic Detection) 17-444

BrdU and Ki-67 QCI / HCA Assay Kit HCS213

MILLIPLEX® map Human Multi-Pathway Signaling Kit 48-680

guava® Cell Cycle Reagent Propidium Iodide Solution 4500-0220

Best-selling Individual Inhibitors of Signaling Pathways

Antibodies

Assays

Key Related Products

To view our complete Inhibitor Resource page, including product listings, technical tips, and our substructure searchable

database, please visit www.emdbiosciences.com/Inhibitors.

26

Related Research Support ToolsEnsure that your discoveries have the highest impact and biological relevance by starting with quality cell and tissue samples. EMD Millipore’s sterile filtration tools, cultureware, and automated cell counting system enable robust, uniform, contamination-free cell and tissue culture.

Sterile FiltrationEliminating contaminants from your cell growth media and additives is absolutely crucial to preserving the integrity and accuracy

of your cell cultures. EMD Millipore’s comprehensive line of sterile filtration tools have been specifically designed to address these

needs and to ensure the reproducibility of your cancer research.

Vacuum-Driven Sterile FiltersStericup® filter devices combine a filter unit with a receiver flask and cap for processing and storage.

Description Membrane/Application Pore Size (µm)Funnel Capacity (mL) Receiver Bottle Catalogue No.

Stericup®-GP Filter Units Millipore Express® PLUS (PES) / fast filtration of tissue culture media and buffers

0.22 500 500 mL SCGPU05RE

Cultureware

Description No. of Layers Total Surface Area (cm2) Qty/Pk Catalogue No.

Millicell® HY FlaskSTEM CELL TESTED

3 600 16 PFHYS0616

5 1000 8 PFHYS1008

Description System Components Membrane/Pore Size Qty/Pk Catalogue No.

Millicell®-96 cell culture insert plates

96-well cell culture plate, single-well feeder tray and lid Isopore (Polycarbonate) 5 PSHT004R5

96-well cell culture plate, 96-well receiver tray and lid Isopore (Polycarbonate) 5 PSHT004S5

Description Qty/Pk Catalogue No.Millicell® EZ slide (4-well glass) 16 PFHYS0616

8 PFHYS1008

Millicell® EZ slide (8-well glass) 16 PEZGS0816

96 PEZGS0896

Millicell® EZ slide Microscope Slide Holder 1 PEZXMSH01

Millicell® Membrane-Based Cell CultureFor more relevant cell-based assays, try growing your cells on EMD Millipore’s membrane-based cell culture products. The

optimized membranes result in cells with structure and function that more closely mimic what occurs in vivo. Obtain high

quality results for screening, cell signaling assays, proliferation studies, and more.

Millicell® HY (High-Yield) Cell Culture FlasksSimplify your cell culture. Obtain robust cell growth for your next big experiment by using

Millicell® HY multilayer flasks to save time, incubator space, reagents, and money.

Millicell® EZ slides

Use the Millicell® EZ slide to culture, fix, stain and view your cells all in one device. There’s no need to tediously move cover slips from your

culture dish to a slide. Leave the wells intact to fix and stain and acquire data simply and quickly with Millicell® EZ slides.

27

Use with InhibitorSelect™ EGFR Signaling Pathway Inhibitor Panel Phosphorylated EGFR pathway proteins, as well as total EGFR, were simultaneously detected in A431 cells treated with 1000, 333, 111, 37,12.3 and 0 ng/mL EGF, using the MILLIPLEX® map EpiQuant™ EGFR Pathway Magnetic Bead Panel (Catalogue No. MPEQMAG-110K). Lysates were collected after 5 minutes EGF stimulation. Values are internally normalized utilizing the TAFII68 loading control.

[EGF] (ng/ mL)

[Ana

lyte

] (pM

)

EpiQuant EGFR Panel: EGF Dose Response

EGFR (pY1110/1125)ERK1/2 (pY204/187)

1

10

100

1,000

10,000

1 10 100 1,000 10,000

EGFR (pY1069/1092)

Shc (pY349/350)ErbB3 (pY1197/1307)

Versatility in a Complete Package.New MILLIPLEX® map EpiQuant™ Assays for Quantitative Cell Signaling Analysis

Quantitating the change in signaling in response to small

molecules can elucidate the relationship between a signal

and its ultimate output. EpiQuant™ technology, the most

advanced platform for cell signaling analysis, is the only

assay that enables absolute quantitation of the phospho-

rylation status of signal transduction proteins, providing

a thorough understanding of EGFR and other signaling

pathways.

Discover the most versatile multiplex platform for your Cell Signaling research.

The MILLIPLEX® map Advantage:

• The largest portfolio of magnetic bead assays— compatible with MAGPIX®, the latest, small- footprint instrument for the Luminex® platform

• Quality and lot-to-lot consistency built into every kit

• Flexible—use with any magnetic bead washer or vacuum manifold/filter plate

• Convenient—packaged in a single kit with a single catalogue number

EMD Millipore, the M mark, K-LISA, InSolution, and InhibitorSelect are trademarks, and Calbiochem and StemSelect are registered trademarks of Merck, KGaA, Darmstadt, Germany.Cell Cycle Stop, ChemiScreen, easyCyte, InCyte, Scepter, DART, MapMate, AXIS, QCM, ECMatrix, Milli-Mark, STEMCCA, and FlowCellect are trademarks of Millipore Corporation.MILLIPLEX, Millicell, Stericup, ApopTag, guava, and ViaCount are registered trademarks of Millipore Corporation.Alexa Fluor is a registered trademark and MitoSOX is a trademark of Life Technologies, Inc.Dell and Latitude are registered trademarks of Dell, Inc. Intel is a registered trademark of Intel Corporation.Lit No. PB3337EN00 LS-SBU-11-04938 Printed in the USA 11/2011 © 2011 Millipore Corporation. All rights reserved.

www.emdmillipore.com

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