YOUR SOURCE for: Biomarker Immunoassays Transforming the Process of Discovery

www.millipore.com/bmia_immuno

• Choose from over 90 individual

cytokine, chemokine and soluble cytokine receptor targets for ultimate assay flexibility

• Detect low levels of inflammation with Millipore’s MILLIPLEX® MAP High Sensitivity Human Cytokine Panel

• Use Millipore’s multi-species TGFβ1 single plex, or our TGFβ 3-plex to investigate inflammation, angiogenesis, normal development and cancer

• Elucidate the role of cytokine biomarkers involved in sepsis and in the apoptotic pathway

• Use our unique skin multiplex panel for the simultaneous quantitation of five important biomarkers for the functional study of the dermis and its sublayers

• Choose from human, mouse and rat multiplex isotyping products to quantify multiple immunoglobulin levels consistently in serum or cell culture supernatant in a single well, saving time and reagents

METABOLISM (Pg. 6)• Choose from the broadest,

continuously expanding offerings of RIA, ELISA and multiplex analytes

• Study metabolic and gut hormones, adipocytes and cardiovascular disease with comprehensive MILLIPLEX MAP

panels.• Correlate levels of endocrine

biomarkers involved in obesity and type 2 diabetes with thyroid and metabolic hormones using Millipore’s MILLIPLEX MAP panels

• Measure multiplexed bone metabolism biomarkers in humans, mice or rats to study osteogenesis, bone resorption and bone metastasis

• Detect levels of pituitary and stress hormones using MILLIPLEX map panels

NEUROSCIENCE (Pg. 11)• Millipore is the leader in multiplex

kits for the study of neuroscience, including panels for neurodegeneration and other neurological disorders.

• Investigate the pathogenesis of diseases such as Alzheimer’s, Parkinson’s and Lewy Body Disease with MILLIPLEX MAP neurodegenerative

panels

TOXICITY (Pg. 12)• Use MILLIPLEX MAP renal toxicity

panels to determine the effects of drug and environmental compounds using animal and human models

• Determine compound safety early in the drug discovery and development process using Millipore’s cell-line-

based ToxReporter™ ELISAs

CANCER (Pg. 14)• Use our soluble cancer biomarker

panel for studying the biochemical mechanisms of, and the body’s response to, tumor development

• Detect levels of activated intracellular proteins involved in apoptosis using Millipore’s MILLIPLEX MAP Apoptosis kit, as well as apoptosis-relevant MAPmates™

CELL SIGNALING (Pg. 16)MILLIPLEX map Multiplex Kits• Create a custom kit using any of the

expanding line of 90 phosphorylated active or total MAPmates

• Choose a pre-configured kit to study multi-pathway signaling, MAPK, Src family kinases, T-cell receptor activation, or the Akt/mTOR pathway

• Use as little as 25 μL or less of cell lysate for cost effective analysis

MILLIPLEX map EpiQuant™

Multiplex Kits (Pg. 19)• Assess the functional states of cells

by profiling phosphorylation changes across signaling pathways and cell phenotypes

• Make site-specific, quantitative measurements with picomolar sensitivity

• Measure total and multiple specific phosphorylated sites in the same well

• Design kits by choosing from large panels.

Lead Discovery SignalProfiler™ Service (Pg. 25)

• Inspection of common intracellular signal transduction cascades and networks

• Cell-based assays for compound efficacy and liability

• Run by experts in compound

selectivity-profiling

LUMINEX® INSTRUMENTATION AND MILLIPLEX ANALYST (Pg. 26)• A highly flexible, xMAP®-based platform

for combining and screening analytes• Efficient quantitation of up to 100

analytes in a single 25 μL sample• Generates information with speed, low

sample volume and economy• Easy-to-use software packages for

better data acquisition and analysis

BIOPHARMA AND BIOMARKER SERVICES (PG. 29)• Millipore has been performing

biomarker studies for over 25 years• Assay services are available for RIAs,

ELISAs, IRMAs and multiplex assays using Millipore kits or any commercially available assay

• Quality control, reproducible results, and confidentiality are of paramount importance to our team

• We offer proven bioanalytical services for biomarkers from research-grade analysis to fully GXP-compliant assay validation and sample analysis. A “fit-for-purpose” validation strategy is implemented to customize the assay to meet your specific regulatory needs

INFLAMMATION/IMMUNOLOGY (Pg. 2)

What’s Inside:

YOUR SOURCE FOR FASTER PROTEIN BIOMARKER ANALYSISIn today’s exciting world of biomarker research, advances are

unfolding rapidly, providing new hope for treating many

diseases. The need to gain quickly an understanding of

protein-receptor interactions, intracellular signaling pathways

and extracellular markers relevant to many biological

processes is greater than ever.

At Millipore, we understand the challenges you face. That’s

why we offer one of the largest portfolios of biomarker

assays—including RIAs, ELISAs and multiplexed assays—for

disease states such as metabolic, neurodegenerative and

cardiovascular diseases, cancer, autoimmune disease and

more. By responding to the needs brought on by new

discoveries, Millipore provides you with access to some of the

most innovative assays and breakthrough platform

technologies in the marketplace. Our kits and reagents

provide you with consistent, high-quality products for faster,

more efficient research, drug discovery and drug development.

TRUST OUR GOLD-STANDARD RIAS AND ELISAS• The largest offering of ELISAs and RIAs supporting research in

metabolic disease

• Built on decades of experience in developing antibodies for

metabolic disorders and cell signaling pathways

• Cost effective when studying one or two analytes

MILLIPLEX maP MAGNETIC AND NON-MAGNETIC MULTIPLEX ASSAY PORTFOLIOMillipore is in the process of converting our MILLIPLEX MAP

multiplex portfolio from a non-magnetic to a magnetic bead

format. Both non-magnetic and magnetic capture beads are

fluorescently dyed polymer beads with specific antibodies

attached to surface carboxyl groups. Consequently, the

magnetic beads share the same surface chemistry, but also

contain encapsulated magnetite. Because surface chemistries

are equivalent to non-magnetic beads, magnetic beads exhibit

equivalent ligand binding affinity and specificity, enabling easy

method transfer and assay setup.

Advantages of the magnetic bead format include:• Fewer issues with clogged wells and leaky plates

• Walk-away washing option for increased convenience

• High-throughput capability

• Flexible—can be washed automatically with a magnetic bead

washer or manually with a handheld magnetic bead washer

or with filter plates

Throughout this brochure, catalogue numbers are bold, the

dark green font denotes our magnetic bead format, while a light green font will indicate our non-magnetic bead format.

MILLIPLEX MAP provides a whole new level of tools and services to help you do your best work.

• The broadest selection of analytes

across a wider range of disease states

and species compared to any other

Luminex partner

• All the components and reagents you

need to detect multiple analytes

simultaneously in a small sample size

(10-50 μL)—all in a single kit, using a

single catalogue number

• The ability to select a pre-mixed kit or

choose analytes within most of our

panels to design a custom kit

• Quality controls provided to qualify

assay performance

• Analytically validated panels that yield

consistent analyte profiles irrespective

of plex size

• Standardized standard curve ranges

across analytes and lots to ensure

lot-to-lot consistency

• Panels meet stringent manufacturing

criteria to ensure batch-to-batch

reproducibility

• Environmentally friendly packaging: our

boxes are 100% recyclable to reduce

ecological footprints

• Consolidated packaging available for

larger orders to reduce storage needs

1

2

The immune system is an organized, complex network of biological struc-

tures and processes that protect against disease. For example, cytokines

and chemokines, proteins of the inflammation pathway, mediate interactions

between cells, regulating target immune cell responses. These responses

result in inflammation, in which the immune system attempts to eradicate

foreign antigens and begin the healing process. Consequently, the inflam-

matory process plays a key protective role in immunity. Millipore’s

MILLIPLEX MAP Cytokine/Chemokine Panels, including our Human High

Sensitivity Cytokine panel, enable you to investigate simultaneously the

modulation and expression of many cytokines in a variety of inflammatory

diseases. To address the requests and needs of many of our customers,

Millipore will soon offer human, mouse and rat panels in both magnetic and

non-magnetic bead formats.

WITH OVER 90 CYTOKINE/CHEMOKINE ANALYTES AND SOLUBLE CYTOKINE RECEPTORS, MILLIPORE IS YOUR SOURCE FOR CHOICE AND FLEXIBILITY• Rely on our analytically validated panels, to ensure consistent analyte

performance

• Select a pre-mixed kit or choose analytes from a validated panel to

design a custom kit

• Human Panel I: 42 analytes; Panel II: 23 analytes; Panel III: 11 analytes

• Human High Sensitivity: 13 analytes

• Non-Human Primate: 23 analytes

• Mouse Panel I: 32 analytes; Panel II: 12 analytes; Panel III: 6 analytes

• Rat: 23 analytes

• Canine: 14 analytes

Fixed Ranges for Standard Curves and Analytes Result in Consistency from Lot to Lot

Millipore’s MILLIPLEX MAP panels offer you industry-leading choices and quality for measuring cytokines, chemokines, and analytes for studying immunogenicity, apoptosis, sepsis and TGFβ expression. Our assay development scientists were the first to produce commercial kits for the Luminex platform. Today, this forward-thinking spirit continues with the most versatile system available for inflammation/immunology research.

INFLAMMATION/IMMUNOLOGYYour Source for More Flexibility and Choice

25,000

20,000

15,000

10,000

5,000

010-4 10-3

Concentration of Each Cytokine(ng/mL)

hIL-6hIL-8hG-CSFhIL-1αhIL-1βhTNF-α

MFI

10-2 10-1 100 101 102 103

15,000

12,500

10,000

7,500

5,000

2,500

010-4 10-3

Concentration of Each Cytokine(ng/mL)

MultiplexSingle-Plex

hG-CSF

MFI

10-2 10-1 100 101 102 103

25,000

20,000

15,000

10,000

5,000

010-4 10-3

Concentration of Each Cytokine(ng/mL)

hIL-6hIL-8hG-CSFhIL-1αhIL-1βhTNF-α

MFI

10-2 10-1 100 101 102 103

15,000

12,500

10,000

7,500

5,000

2,500

010-4 10-3

Concentration of Each Cytokine(ng/mL)

MultiplexSingle-Plex

hG-CSF

MFI

10-2 10-1 100 101 102 103

MILLIPLEX maP validated panels yield consistent performance irrespective of plex size

Multiplex Cytokines Multiplex vs. Single-Plex

Experience industry-leading choice and quality, while improving your productivity

Cytokines/Chemokines

HUMAN PANEL I(Cat. No. HCYTOMAG-60K)(Cat. No. MPXHCYTO-60K)

EGFEotaxinFGF-2Flt-3 LigandFractalkineG-CSFGM-CSFGROIFNa2IFNgIL-1aIL-1βIL-1raIL-2IL-3IL-4IL-5IL-6IL-7IL-8IL-9

IL-10IL-12 (p40)IL-12 (p70)IL-13IL-15IL-17IP-10MCP-1MCP-3/CCL7MDC/CCL22MIP-1aMIP-1βPDGF-AAPDGF-AB/BBRANTESsCD40LsIL-2RaTGFaTNFaTNFβVEGF

HUMAN PANEL II(Cat. No. HCYP2MAG-62K)(Cat. No. MPXHCYP2-62K)

6Ckine/CCL21BCA-1/CXCL13CTACK/CCL27ENA-78/CXCL5Eotaxin-2/CCL24/ MPIF-2Eotaxin-3/CCL26I-309/CCL1IL-16IL-20IL-21IL-23IL-28a

IL-33/NF-HEV (mature)LIFMCP-2MCP-4MIP-1d (MIP-5/ CCL15)SCFSDF-1a+β/ CXCL12TARC/CCL17TPOTRAIL/TNFSF10TSLP

HUMAN PANEL III(Cat. No. HCP3MAG-74K)(Cat. No. MPXHCYP3-63K)

GCP-2/CXCL6/LIXHCC-1/CCL14aIL-11IL-29I-TAC/CXCL11LymphotactinM-CSFMIG/CXCL9MIP-3a/CCL20MIP-3β/CCL19NAP-2/CXCL7

HUMAN HIGH SENSITIVITY(Cat. No. HSCYTMAG-60SK)(Cat. No. HSCYTO-60SK)

GM-CSFIFNgIL-1βIL-2IL-4IL-5IL-6IL-7IL-8IL-10IL-12 (p70)IL-13TNFa

NON-HUMAN PRIMATE(Cat. No. MPXPRCYTO-40K)

G-CSFGM-CSFIFNgIL-1βIL-1raIL-2IL-4IL-5IL-6IL-8IL-10IL-12/23 (p40)

IL-13IL-15IL-17IL-18MCP-1MIP-1aMIP-1βsCD40LTGFaTNFaVEGF

MOUSE PANEL I(Cat. No. MCYTOMAG-70K)(Cat. No. MPXMCYTO-70K)

Eotaxin G-CSFGM-CSFIFNgIL-1aIL-1βIL-2IL-3IL-4IL-5IL-6IL-7IL-9IL-10IL-12 (p40)IL-12 (p70)

IL-13IL-15IL-17IP-10KCLIFLIXMCP-1M-CSFMIGMIP-1aMIP-1βMIP-2RANTESTNFaVEGF

MOUSE PANEL II(Cat. No. MCYP2MAG-73K)(Cat. No. MPXMCYP2-73K)

EPOExodus-2FractalkineIL-16IL-21IL-22IL-25IL-28BMCP-5MIP-1aMIP-1βTARC

MOUSE PANEL III(Cat. No. MPXMCYP3-74K)

IL-20IL-23IL-27IL-33MDCTIMP-1

RAT(Cat. No. RCYT0MAG-80K)(Cat. No. RCYTO-80K)

EotaxinG-CSFGM-CSFGRO/KCIFNgIL-1aIL-1βIL-2IL-4IL-5IL-6IL-10IL-12 (p70)IL-13IL-17IL-18IP-10LeptinMCP-1MIP-1aRANTESTNFaVEGF

CANINE(Cat. No. CCYTO-90K)

GM-CSFIFNgIL-2IL-4IL-6IL-7IL-8IL-10IL-15IL-18IP-10KC-likeMCP-1TNFa

3

INFLAMMATION/IMMUNOLOGY

NOTE: Catalogue numbers appearing in dark green font denote our magnetic bead format.

THE POWER OF MILLIPORE’S SOLUBLE CYTOKINE RECEPTOR PANELWhile cytokines play a fundamental role in the pathogenesis of

many diseases, the discovery that soluble cytokine receptors

are involved in regulating excessive inflammatory responses

and modulating immune events has stimulated significant

research interest in their potential role as immunotherapeutic

agents. Responding to this need, Millipore provides the first

Human and Mouse Soluble Cytokine Receptor Multiplex Panels

using the Luminex xMAP technology.

Soluble Cytokine Receptor Panels for the Luminex xMAP Platform

SEPSISMillipore provides important tools to understand better the

role of cytokine biomarkers involved in sepsis and in the

apoptotic pathway. Based on the Luminex xMAP platform, the

MILLIPLEX MAP Human Sepsis Panel assists you in your

research to find an effective treatment for Systemic

Inflammatory Response Syndrome (SIRS).

IMMUNOLOGY ELISAS: CIRCULATING BIOMARKERS

Human Soluble Receptor (Cat. No. HSCR-32K)

sCD30sEGFRsgp130sIL-1RIsIL-1RIIsIL-2RasIL-4RsIL-6RsRAGEsTNF RIsTNF RIIsVEGF R1sVEGF R2sVEGF R3

Human Sepsis Panel 1 (Cat. No. HSEP-63K)

MIFsFassFasLsICAM-1PAI-1 (total)sVCAM-1

Human Sepsis Panel 3

Elastase 2LactotransferrinLipocalin (NGAL)ResistinThrombospondin 1

Human Sepsis Panel 2

Granzyme BHSP70IL-1aIL-8MIP-1aMIP-1βMMP-8

Mouse Soluble Receptor (Cat. No. MSCR-42K)

sCD30sgp130sIL-1RIsIL-1RIIsIL-2RAsIL-4RsIL-6RsRAGEsTNF RIsTNF RIIsVEGF R1sVEGF R2sVEGF R3

4

INFLAMMATION/IMMUNOLOGY

Kit Species Cat. No.

sCD26 Human ECM345

sE-Selectin Human ECM330

Fibronectin (Quantimatrix) Human ECM300

ICAM-3 Human ECM337

SICAM-2 Human ECM346

IFNA a Human CYT271

Laminin Human ECM310

PEGF Human CYT420

sL-Selectin Human ECM331

Human Sepsis Panel

Transforming Growth Factor Multiplex Panels

MULTI-SPECIES TRANSFORMING GROWTH FACTOR β PANELSTGFβ’s involvement in basic biological processes and the op-

posing roles that both the normal and mutated proteins play

in tumor cell proliferation emphasizes the importance of this

cytokine in a variety of research areas. Based on the Luminex

xMAP platform, our MILLIPLEX MAP Multi-Species TGFβ1 Single

Plex and TGFβ 3-Plex (TGFβ1, TGFβ2, TGFβ3) kits have been

optimized for performance and ease of use.

Multi-Species (Cat. No. TGFB-64K-01)

TGFβ1

Multi-Species (Cat. No. TGFB-64K-03)

TGFβ1TGFβ2TGFβ3

NOTE: Catalogue numbers appearing in dark green font denote our magnetic bead format.

ISOTYPINGImmunoglobulin isotyping is essential for characterizing puri-

fied antibodies and the immunogenicity of therapeutic proteins

for monitoring and predicting reactions to biopharmaceuticals.

Millipore’s Human, Mouse and Rat MILLIPLEX map Isotyping kits

enable you to quantify in multiple immunoglubulin levels con-

sistently in serum or cell culture supernatant in a single well,

saving time and reagents compared to ELISAs. Panels in the

magnetic bead format are also customizable, enabling you to

choose the immunoglobulins that best meet your needs.

HUMAN SKIN PANELSkin is the largest organ in the body, responding to damage by

upregulating repair mechanisms and causing skin hyperprolifera-

tion. Consequently, analysis of biochemical components from the

stratum corneum (SC) can be used to evaluate skin irritation, SC

integrity and structure and inflammatory changes in human skin.

Millipore’s MILLIPLEX map Human Skin Panel enables you to analyze

analyte concentrations in skin cell extracts or skin samples col-

lected with a non-invasive tape absorption technique.

Human Isotyping(Cat. No. HGAMMAG-301K) (Cat. No. HGAM-301)

IgAIgG1IgG2IgG3IgG4IgM

Mouse IgE Single Plex(Cat. No. MGAM-302E)

IgE

Mouse Isotyping (Cat. No. MGAM-300)

IgAIgG1IgG2aIgG2bIgG3lgM

Human IgE Single Plex (Cat. No. HGAMMAG-303E)

IgE

Human Skin(Cat. No. SKIN-50K)

CortisolFibronectin Human Serum AlbuminInvolucrinLPSKeratin-6Keratin-1,10

INFLAMMATION/IMMUNOLOGY

0

1

10

100

1,000

10,000

100,000

EGF

Eotaxin

FGF-2

FIt-3L

Fractalkine

G-CSF

GM-CSF

GRO

IFNα2

IFNγ

IL-1α

IL-1β

IL-1ra

IL-2

IL-3

IL-4

IL-5

IL-6

IL-7

IL-8

IL-9

IL-10

IL-12p40

IL-12p70

IL-13

IL-15

IL-17

IP-10

MCP-1

MCP-3

MDC

MIP-1α

MIP-1β

sCD40L

sIL-2R

α

TGFα

TNFα

TNFβ

VEGF

PDGF-AA

PDGF-AB/BB

RANTES

Con

cent

rati

on (pg

/mL) Normal

Sepsis

Cytokine and Chemokine Concentrations in Normal and Septic Human Serum Samples

The MILLIPLEX MAP Cytokine/Chemokine 42-Premixed panel was used to determine cytokine and chemokine concentrations (pg/mL) in serum samples of both normal (n=20) and sepsis (n=10) patients. The results of the assay are shown above.

5

Isotyping Multiplex Panels

6

METABOLIC DISEASE, CARDIOVASCULAR DISEASE AND THE ENDOCRINE SYSTEMWith the rise in incidence of atherosclerosis, coronary heart disease, type 2

diabetes and kidney disease, Metabolic Syndrome has received increased

attention from the medical community in recent years. That’s why research-

ers everywhere rely on Millipore. With RIAs, ELISAs and multiplex detection

kits, Millipore leads the drug discovery industry by providing the greatest

variety of analytically validated assays and platforms for the study of

metabolic syndrome, helping you better understand its causes.

FAT

HYPOTHALAMUS

PANCREAS

GUT

NPY

MCH

AgRP

Orexin

GLP-1

GIPGLP-2

GRP

PYYGastrinOxyntomodulin

VIP

CCK

ObestatinProinsulinC-PeptideAmylin

GlucagonPPSomatostatin

IL-6IL-8IL-10TNFαMCP-1MIFPAI-1ASP

Adipsin

1

n

CortisolHGFNGFVEGFVisfatinVaspinRBP4Chemerin

Available from Millipore

Adiponectin

Resistin

Ghrelin

PYY3-36

Insulin Myostatin

Musclin

MUSCLE

Food Intake

EnergyExpediture

BodyWeight

Leptin

METABOLISMYour Source for Comprehensive Analysis from Every Angle

DIABETES AND OBESITY BIOMARKERS

We’re constantly expanding and enhancing our already broad portfolio of RIAs, ELISAs and MILLIPLEX map panels. Whether you’re a researcher who needs to explore highly intricate protein combinations in greater detail or a lab technician who needs to test samples simultaneously and interpret data, Millipore can help move your mission-critical work forward.

7

The Greatest Variety of Highly Validated Assays and Platforms for the Study of Metabolic Syndrome

Kit Species Catalogue No.

Adiponectin Human HADP-61HK

Adiponectin Mouse MADP-60HK

C-Peptide Canine CCP-24HK

C-Peptide Human HCP-20K

C-Peptide Porcine PCP-22K

C-Peptide Rat RCP-21K

Ghrelin (active) Human GHRA-88HK

Ghrelin (total) Human GHRT-89HK

Glucagon GL-32K

GLP-1 (active) GLP1A-35HK

GLP-1 (total) GLP1T-36HK

Growth Hormone Porcine/Canine PGH-46HK

Growth Hormone Rat RGH-45HK

Insulin Lispro LPI-16K

Insulin Porcine PI-12K

Insulin Rat RI-13K

Insulin Sensitive Rat SRI-13K

Insulin Ultra Sensitive Human HI-11K

Insulin Specific Human HI-14K

Leptin Human PL-81K

Leptin Mouse ML-82K

Leptin Multi Species XL-85K

Leptin Primate HL-81K

Leptin Rat RL-83K

Pancreatic Polypeptide Rat RPP-41HK

Proinsulin Human HPI-15K

PYY (total) Human PYYT-66HK

PYY Rat/Mouse RMPYY-68HK

PYY (3-36) Specific Human PYY-67HK

Kit Species Catalogue No.

Adiponectin• Human EZHADP-61K

Adiponectin – High Molecular Weight•

Human EZHMWA-64K

Adiponectin• Mouse EZMADP-60K

Adiponectin• Canine EZCADP63K

Adiponectin• Rat EZRADP-62K

Adipsin• Mouse EZMADPSN-22K

AGF (ANGPTL6) Human EZHAGF-28K

AGP Mouse EZMAGP-23K

Amylin (active) Human EZHA-52K

Amylin (total) Human EZHAT-51K

Apo IV Human EZHAPOA4-73K

Apo V Human EZHAPOAV71K

BDNF Human/Rat CYT306

Chemerin Human EZHCMRN-57K

C-Peptide• Human EZHCP-20K

C-Peptide 2• Rat/Mouse EZMCP2-21K

CRP Canine CYT296

CRP Rat CYT294

CRP Human CYT298

FGF-21 Human EZHFGF21-19K

FGF-21 Rat/Mouse EZRMFGF21-26K

FGF-23 Human EZHFGF23-32K

Ghrelin (active)• Human EZGRA-88K

Ghrelin (active)• Rat/Mouse EZRGA-90K

Ghrelin (total)• Human EZGRT-89K

Ghrelin (total)• Rat/Mouse EZRGRT-91K

GIP (total)• Human EZHGIP-54K

GIP (total)• Rat/Mouse EZRMGIP-55K

GLP-1 (active) Multi-Species EGLP-35K

GLP-1 (total) Multi-Species EZGLP1T-36K

GLP-2 (total) Human/Rat EZGLP2-37K

Growth Hormone• Rat/Mouse EZRMGH-45K

Insulin• Human EZHI-14K

Insulin• Human (Animal Serum Free)

EZHIASF-14K

Insulin• Rat/Mouse EZRMI-13K

Proinsulin (Intact)• Human EZHIPI-17K

Proinsulin (total)• Human EZHPI-15K

Leptin• Canine EZCL-31K

Leptin• Mouse EZML-82K

Leptin• Rat EZRL-83K

Leptin “Dual Range”• Human EZHL-80SK

Pancreatic Polypeptide• Human EZHPP-40K

Procollagen Type IIA N-Propeptide† (PIIANP)•

Human EZPIIANP-53K

PYY (total)• Human EZHPYYT-66K

RBP4 Human EZHRBP4-18K

Resistin• Human EZHR-95K

Serum Amyloid A-3 (SAA-3) Mouse EZSAA3-12K

RIAs ELISAs

† U.S. Patent #:5,541,066:5,780,240 • Strip Plate Format

METABOLISM

8

• Choose from the broadest selection of ELISAs, RIAs and

multiplex detection kits, in the broadest range of species

available.

• Select from our continuously expanding menu of MILLIPLEX

MAP multiplex panels for the study of metabolic disease

markers in a broad range of species.

• Customize each multiplex panel by choosing those analytes

within a panel that meet your multiplexing needs.

• Generate information with speed, low sample volume and

economy using our endocrine, cardiovascular disease and

gut hormone metabolic disease multiplex panels.

Endocrine Multiplex Kits

HUMAN ENDOCRINE (Cat. No. HENDO-65K)

Amylin (active)Amylin (total)C-PeptideGLP-1 (active)

GlucagonInsulinLeptin

MOUSE ENDOCRINE (Cat. No. MENDO-75K)

Amylin (active)GLP-1 (active)Glucagon

InsulinLeptin

RAT ENDOCRINE (Cat. No. RENDO-85K)

Amylin (active) GLP-1 (active) Glucagon

InsulinLeptin

HUMAN IGF BINDING PROTEIN (IGFBP) PANEL (Cat. No. HIGFBP-53K)

IGFBP1IGFBP2IGFBP3IGFBP4

IGFBP5IGFBP6IGFBP7

MOUSE IGF BINDING (IGFBP) PANEL(Cat. No. MIGFBPMAG-43K)(Cat. No. MIGFBP-43K)

IGFBP1IGFBP2IGFBP3

IGFBP5IGFBP6IGFBP7

HUMAN BRAIN-DERIVED PROTEIN PANEL (Cat. No. HBDP-33K)

ACTHAGRPBDNFCNTFFSH

GHLHPRLTSH

RAT THYROID SINGLE PLEX

TSH

RAT THYROID TWO PLEX (Cat. No. RTHY-30K-T3T4)

T3 T4

RAT THYROID THREE PLEX (Cat. No. RTHY-30K-03)

T3T4

TSH

MULTI-SPECIES STEROID/THY-ROID HORMONE(Cat. No. STTHMAG-21K)

CortisolEstradiolT3

T4ProgesteroneTestosterone

SINGLE PLEX KITS

Adiponectin (Mouse) Adiponectin (Rat) IGF-1 (Human) IGF-2 (Human) IGF-1 (Rat/Mouse)

Gut Hormone Multiplex Kits

HUMAN GUT HORMONE (Cat. No. HGT-68K)

Amylin (active)Amylin (total)Ghrelin (active)GIP (total)GLP-1 (active)InsulinLeptinPancreatic PolypeptidePYY (total)

RAT GUT HORMONE(Cat. No. RGT-88K)

Amylin (active)Ghrelin (active)GIP (total)GLP-1 (active)InsulinLeptinPancreatic PolypeptidePYY (total)

Metabolic Multiplex Kits

HUMAN METABOLIC DISEASE (Cat. No. HMHMAG-34K) (Cat. No. HMH-34K)

Amylin (active)Amylin (total)C-PeptideGhrelin (active)GIP (total)GLP-1 (active)GlucagonIL-6

InsulinLeptinMCP-1PancreaticPolypeptidePYY (total)TNFa

MOUSE METABOLIC DISEASE (Cat. No. MMHMAG-44K)

Amylin (active)C-PeptideGhrelin (active)GIP (total)GLP-1 (active)Glucagon L-6

InsulinLeptinMCP-1PancreaticPolypeptidePYY (total) ResistinTNFa

RAT METABOLIC DISEASE (Cat. No. RMHMAG-84K)

Amylin (active)C-Peptide 2Ghrelin (active)GIP (total)GLP-1 (active)GlucagonIL-6 Insulin

LeptinMCP-1Pancreatic Polypeptide (one analyte)PYY (total)TNFa

HUMAN LIVER PROTEIN (Cat. No. HLPPMAG-57K)(Cat. No. HLPP-57K)

AFPANGPTL3ANGPTL4ANGPTL6FABP1

FGF-19FGF-21FGF-23HGF

HUMAN ADIPOKINE PANEL A (serum/plasma samples) (Cat. No. HADK1-61K-A)

AdiponectinPAI-1 (active)

PAI-1 (total)Resistin

HUMAN ADIPOKINE PANEL B (serum/plasma samples) (Cat. No. HADK2-61K-B)

HGFIL-1βIL-6IL-8Insulin

LeptinMCP-1NGFTNFa

MOUSE ADIPOKINE (serum/plasma samples) (Cat. No. MADPK-71K)

IL-6InsulinLeptinMCP-1

PAI-1 (total)ResistinTNFa

RAT ADIPOKINE (serum/plasma samples) (Cat. No. RADPK-81K)

IL-1βIL-6InsulinLeptin

MCP-1PAI-1 (active)PAI-1 (total)TNFa

CANINE ADIPOKINE (Cat. No. CADPK-91K)

Adiponectin Resistin

HUMAN ADIPOCYTE (for cell culture samples) (Cat. No. HADCYT-61K)

AdiponectinHGFIL-1βIL-6IL-8Leptin

MCP-1NGFPAI-1 (active)PAI-1 (total)ResistinTNFa

METABOLISM

MOUSE GUT HORMONE (Cat. No. MGT-78K)

Amylin (active)Ghrelin (active)GIP (total)GLP-1 (active)InsulinLeptinPancreatic PolypeptidePYY (total)

CANINE GUT HORMONE (Cat. No. CGTMAG-98K)(Cat. No. CGT-98K)

Amylin (total)Ghrelin (active)GIP (total)GLP-1 (active)GlucagonInsulinPancreatic PolypeptidePYY (total)

YOUR SOURCE FOR UNLOCKING THE MYSTERIES OF METABOLIC SYNDROME

NOTE: Catalogue numbers appearing in dark green font denote our magnetic bead format.

9

METABOLISM

Cardiovascular Multiplex Kits

HUMAN CVD1 (Cat. No. HCVD1-67AK)

AdiponectinMMP-9MPOPAI-1 (total)sE-SelectinslCAM-1sVCAM-1

HUMAN CVD2 (acute phase proteins)(Cat. No. HCVD2-67BK)

CRPSerum Amyloid ASerum Amyloid P

HUMAN CVD3 (Cat. No. HCVD3-67CK)

IFNgIL-1βIL-6IL-8IL-10MCP-1NT-proBNPTNFaVEGF

SINGLE PLEX HUMAN

FibrinogenHaptoglobin

HUMAN APOLIPOPROTEINS (Cat. No. APO-62K)

Apo AIApo AIIApo BApo CIIApo CIIIApo E

MOUSE CVD1 (Cat. No. MCVD1-77AK)

MMP-9PAI-1 (total)sE-SelectinslCAM-1sVCAM-1

MOUSE CVD2 (Cat. No. MCVD277BK)

AdiponectinApo AIApo EFibrinogen

RAT CVD1 (Cat. No. RPTMAG-86K)(Cat. No. RCVD1-89K)

IL-6BNPMCP-1MPOPAI-1TIMP-1

TNFaTroponin ITroponin TVEGFvon Willebrand Factor

RAT CVD2 (Cat. No. RCVD2-89K)

sE-SelectinsICAM-1

RAT CVD3 (Cat. No. RCVD3-89K)

AdiponectinFibrinogen

SINGLE PLEX RAT / MOUSE

CRP

Pituitary Hormones Multiplex Panels

PITUITARY HORMONESCalled the “master gland” because it controls many other

endocrine glands, the pituitary gland secretes a number of key

hormones that play important roles in the regulation of

metabolism, growth, and reproduction. For example, these

hormones are responsible for the stimulation of the:

• Adrenal gland (ACTH)

• Thyroid gland (TSH)

• Ovaries and testes (FSH, LH)

• Breast milk production (prolactin)

• Blood pressure

• Amount of water excreted by the kidneys

In turn, the pituitary gland is connected to and controlled by

the hypothalamus, which determines the amount of stimulation

the pituitary’s target glands need by detecting their hormone

levels.

CANINE PITUITARY (Cat. No. CPTMAG-96K )

ACTHBDNFFSHGH

LHProlactinTSH

HUMAN BRAIN-DERIVED PROTEIN PANEL (Cat. No. HBDP-33K)

ACTHAGRPBDNF CNTF FSHGH LHProlactinTSH

HUMAN PITUITARY (Cat. No. HPT-66K)

ACTHFSHGHLHProlactinTSH

RAT PITUITARY (Cat. No. RPTMAG-86K) (Cat. No. RPT86K)

ACTHBDNFFSHGHLHProlactinTSH

Stress Hormones Multiplex PanelsSTRESS HORMONESStress is defined as the disruption of homeostasis through

physical and/or psychological stimuli (stressors). Physiologically

the neuroendocrine system, which includes the hypothalamus,

pituitary and adrenal glands, plays a critical role in response to

stress. Activated by corticotrophin-releasing hormone (CRH)

and arginine-vasopressin (AV), the pituitary gland releases

ACTH, which results in the secretion of cortisol /

corticosterone and other glucocorticoids from the adrenal

glands. These corticoids involve the whole body in the stress

response and are finally involved in the termination of the

stress response by a negative feedback loop to the

hypothalamus.

RAT STRESS HORMONE(Cat. No. RSH69K)

ACTHCorticosteroneMelatonin

METABOLISM

10

BONE METABOLISMBiochemical markers of bone metabolism play a significant role

in the assessment of bone diseases such as osteoporosis,

arthritis, chronic inflammatory disorders and cancers. That’s

why we provide the widest range of MILLIPLEX MAP panels for

your drug discovery and research efforts, including panels for

measurement of multiple bone metabolism markers in serum,

plasma or tissue culture supernatant samples. Using these

powerful tools, you’ll advance your study of the biological

functions of these biomarkers and their pathological roles.

Millipore’s Luminex xMAP-based multiplexed bone metabolism

panels are available for the simultaneous measurement of

multiple bone metabolism markers in human, mice or rats.

Each multi-analyte panel is fully customizable to meet your

research requirements.

HUMAN PANEL 1A (serum/plasma samples)(Cat. No. HBN1A-51K)

ACTHInsulinLeptinOPG

OsteocalcinOsteopontinPTH

HUMAN PANEL 1B (Tissue Culture) (Cat. No. HBN1B-51K)

IL-1βIL-6ACTHAdiponectinInsulinLeptinOPG

OsteocalcinOsteopontinPTHTNFa

Single Plex Human

RANKL

MOUSE PANEL 1A (Undiluted serum/plasma samples) (Cat. No. MBN1A-41K)

ACTHIL-6Insulin

LeptinOPGTNFa

MOUSE PANEL 2A (Undiluted serum/plasma samples)(Cat. No. MBN2A-41K)

IL-6ACTHInsulin

LeptinRANKLTNFa

Bone Metabolism Multiplex Kits

MOUSE PANEL 1B (Tissue culture samples) (Cat. No. MBN1B-41K)

IL-6ACTHInsulinLeptin

OPGOsteocalcinTNFa

MOUSE PANEL 2B (Tissue culture samples)(Cat. No. MBN2B-41K)

IL-6ACTHInsulinLeptin

TNFaOsteocalcinRANKL

SINGLE PLEX MOUSE

OPGOsteocalcin

RANKL

RAT PANEL 1 (Cat. No. RBN1-31K)

ACTHInsulin

LeptinOPG

RAT PANEL 2 (Cat. No. RBN2-31K)

ACTHInsulin

LeptinRANKL

RAT PANEL 3 (Cat. No. RBN3-31K)

OsteocalcinOsteopontinPTH

Single Plex Rat

OPGOsteocalcinRANKL

NOTE: Catalogue numbers appearing in dark green font denote our magnetic bead format.

11

Neuroscience ELISAs

NEUROSCIENCE RESEARCH ELUCIDATES MECHANISMS OF NORMAL DEVELOPMENT, HOMEOSTASIS AND DISEASE STATES SUCH AS:• Alzheimer’s Disease

• Parkinson’s Disease

• Lewy Body Disease

• Amyotrophic Lateral Sclerosis (ALS)

• Other Neurological Disorders

HUMAN NEURODEGENERATIVE DISEASE PANEL 1(Cat. No. HNDG1-36K)

a2-MacroglobulinApo AIApo CIIIApo E

Complement C3Complement Factor HPrealbumin

HUMAN NEURODEGENERATIVE DISEASE PANEL 4(Cat. No. HNDG4-36K)

Aβ 1-40Aβ 1-42GDNF

sRAGES100B

HUMAN NEUROLOGICAL DISORDERS PANEL 1 (Cat. No. HND1MAG-39K)

a-SynucleinBDNFGFAPMBPNEFHNEFL

NGF-βNSEPARK5PARK7Tau (pThr231)Tau (total)

HUMAN NEUROLOGICAL DISORDERS PANEL 2 (Cat. No. HND2MAG-39K)

AGPCP

HPSAP

HUMAN NEUROPEPTIDE(requires sample extraction)(Cat. No. HNP-35K)

a-MSHβ-EndorphinCortisolMelatoninNeurotensinOrexin AOxytocinSubstance P

Neuroscience Multiplex Panels

Neurodegenerative Multiplex Panels

NEURODEGENERATIVE DISEASEUnderstanding neurobiology is fundamental to determining the pathogenesis

of neurodegenerative diseases. Identification of key biomarkers and their

accurate measurement is essential. However, conventional methods,

including RIAs and ELISAs, are not able to measure multiple biomarkers

simultaneously with small sample volume. The Luminex-based MILLIPLEX map

Human Neurodegenerative Panels will enable you to explore the complexities

of the nervous system and the pathobiology of disease.

HUMAN NEURODEGENERATIVE DISEASE PANEL 2(Cat. No. HNDG2-36K)

a1-AntitryspinComplement C4CRP

MIP-4PEDFSAP

HUMAN NEURODEGENERATIVE DISEASE PANEL 3(Cat. No. HNDG3-36K)

BDNFCathepsin DMPOPAI-1 (total)PDGF-AA

PDGF-AB/BBRANTESsICAM-1sNCAMsVCAM-1

NEUROSCIENCEYour Source for Measuring Essential Neuroscience Biomarkers Simultaneously

Neurodegenerative disease is a condition characterized by the deterioration of neurons or their myelin sheath over time in the brain and/or spinal cord. These neurons are responsible for everyday activities such as processing sensory information and controlling movement. Because these cells are not easily regenerated, excessive cumulative damage can lead to age-related diseases such as Alzheimer’s and Parkinson’s disease, as well as other conditions such as amyotrophic lateral sclerosis (ALS) and epilepsy.

Description Species Cat. No.

Amyloid β, Set Human EZHS-SET

Amyloid β40 ELISA Kit Human EZBRAIN-40

Amyloid β42 ELISA Kit Human EZBRAIN-42

Amyloid β40 High Sensitivity ELISA Kit Human EZHS40

Amyloid β42 High Sensitivity ELISA Kit Human EZHS42

a-Synuclein HumanMouse Rat

NS400

Chemikine™ Brain Derived Factor Human Rat

CYT306

ChemiKine Nerve Growth Factor Mouse Rat

CYT304

ChemiKine Neurotrophin 3 Human Rat

CYT302

ChemiKine Pigment Epithelium Derived Factor (PEDF)

Human CYT420

Glial Fibrillary Acidic Protein (GFAP) Human MouseRat

NS830

Neuropeptide Y (NPY) Human EZHNPY-25K

Neuropeptide Y (NPY) MouseRat

EZRMNPY-27K

Neurotrophin 3 ELISA-192 Well Human Rat

CYT302

Nitrotyrosine Multi-Species 17-376

OxyELISA Oxidized Protein Quantitation Kit

Multi-Species S7250

Phosphorylated Neurofilament H ELISA Kit Multi-Species NS170

Serum Amyloid A-3 SAA-3 Mouse EZMSAA3-12K

sICAM-1 Human ECM335

sVCAM-1 Human ECM340

NEUROPEPTIDESNeuropeptides are secreted primarily

from the central and peripheral nervous

systems, exerting a broad spectrum of

biological functions that include the

regulation of metabolism, reproduction

and immunity.

12

KIDNEY TOXICITY BIOMARKERSAbsorption, distribution, metabolism and excretion (ADME) parameters are

critical to all stages of a fully integrated drug development program and are

used to augment the interpretation of toxicological findings. As toxicity

represents the leading cause of lead compound failure, ADMET groups

require more sensitive and more rapid methods for determining organ-

specific damage.

Drug-induced damage to kidney cells, also known as nephrotoxicity, can

result from drug excretion. Traditional methods for determining renal

toxicity have relied on the blood urea nitrogen (BUN) and serum creatinine

levels. These two tests only detect kidney damage a week after it begins to

occur and only indicate that damage has occurred somewhere in the kidney.

This has led the FDA/EMEA to begin approving and then mandating specific

biomarkers as part of clinical trials. These biomarkers include:

• Albumin

• β-2 microglobulin

• Clusterin

• Cystatin C

• KIM-1

• Trefoil factor-3 (TFF3)

• Total protein

With Millipore’s MILLIPLEX MAP Kidney Toxicity Panels, you gain the advan-

tages of speed and consistency, allowing quantitative multiplex detection of

multiple analytes simultaneously, dramatically improving productivity.

RAT KIDNEY TOXICITY PANEL 1(Cat. No. RKTX1-37K)

ClusterinKIM-1Osteopontin

RAT KIDNEY TOXICITY PANEL 2(Cat. No. RKTX2-37K)

Albuminβ-2 MicroglobulinCystatin C

HUMAN KIDNEY TOXICITY PANEL 1 (Serum samples)(Cat. No. HKTX1MAG-38K) (Cat. No. HKTX1-38K)

KIM-1OsteopontinReninTFF3

ToxicityHUMAN KIDNEY TOXICITY PANEL 2 (Serum samples) (Cat. No. HKTX2MAG-38K)(Cat. No. HKTX2-38K)

β-2 MicroglobulinClusterinCystatin C

HUMAN KIDNEY TOXICITY PANEL 3 (Urine samples) (Cat. No. HKTX3MAG-38K)(Cat. No. HKTX3-38K)

KIM-1ReninTFF3

HUMAN KIDNEY TOXICITY PANEL 4 (Urine samples) (Cat. No. HKTX4MAG-38K)(Cat. No. HKTX4-38K)

Albuminβ-2 MicroglobulinClusterinCystatin COsteopontinToxicity

NEURODEGENERATIVE DISEASEUnderstanding neurobiology is fundamental to

determining the pathogenesis of neurodegenera-

tive diseases. Identification of key biomarkers

and their accurate measurement is essential.

However, conventional methods, including RIAs

and ELISAs, are not able to measure multiple

biomarkers simultaneously with small sample

volume. The Luminex-based MILLIPLEX map Human

Neurodegenerative Panels will enable you to

explore the complexities of the nervous system

and the pathology of disease.

TOXICITYYour Source for Accelerating Drug Discovery Research

The greatest bottlenecks in drug discovery and development continue to be compound safety, toxicity and unexpected adverse effects during clinical trials. These are areas where new solutions are required—solutions that identify unsafe compounds and either “fail” these drugs as early as possible in the drug discovery and development process or monitor potential risks and flag compounds as they advance through the pipeline. With Millipore’s Kidney Toxicity and ToxReporter cell-based ELISA panels it is now possible to identify a potential toxicity issue earlier in the drug development process. The result is a better understanding of the structural features that drive toxicity pathways, and the ability to detect early biomarkers of potential toxicity in animal and human models.

NOTE: Catalogue numbers appearing in dark green font denote our magnetic bead format.

13

TOXREPORTER CELL-BASED ELISASGiven the high costs and high rates of drug candidate attrition

during clinical trials, early safety assessment across a range of

targets is critical. You need the right tools to help you

discover everything you can about your drug candidates, so

you can avoid costly errors and late-stage compound failure,

developing compounds that offer the greatest promise of

therapeutic success.

That’s why Millipore, in collaboration with CXR Biosciences,

is pleased to offer the ToxReporter ELISA kit. ToxReporter

ELISA’s unique quantitative cell-based toxicity assay enables

you to analyze many of the common cellular events that lead

to compound failure in the clinic or during clinical development,

including:

• Oxidative stress

• Cellular stress

• DNA damage

ToxReporter ELISA includes in vitro cell lines in which a

series of promoters are used to drive the expression of a

common reporter molecule. If the addition of a compound or

chemical to the cell lines induces a mechanism of toxicity, the

reporter protein is secreted into the tissue culture medium. A

single ELISA has been designed to read the reporter molecule,

for all of the ToxReporter cell lines, independent of the specific

response element.

This non-disruptive assay format enables use of live cells

for experiments, allowing time course measurements from a

single sample. ToxReporter cell lines permit the quantitative

detection of early biomarkers of toxicity and cellular stress.

Sources of stress include compounds being screened for drug

discovery, as well as organic or inorganic molecules potentially

requiring environmental monitoring.

ToxReporter Product Line

ECL-001

p21

ECL-002

AP-1

ECL-003

GRE

ECL-004

HSP70

ECLSP-001

Etoposide

ECLSP-002

TPA

ECLSP-003

Dex

ECLSP-004

CuSO4

ECLSP-005

TNFa

ECL-005

NFkB

ToxReporter Cell Lines

ToxReporter Cell Line Starter Packs

ToxReporter ELISA(Catalogue No. ETR-201K)

• Immunosuppression

• Heavy metal toxicity

ECL-006

HRE

ECL-007

XRE

ECL-008

ARE

ECL-009

Hmox1

ECL-010

p53

ECLSP-006

CoCL2

ECLSP-007

3-MC

ECLSP-008

TBHQ

ECLSP-009

Hemin

ECLSP-010

Etoposide

ToxReporter Cell Lines

ToxReporter Cell Line Starter Packs

ToxReporter Cell Line Starter Packs

TOXICITY

14

HUMAN APOPTOSIS 3-PLEX (Cat. No. 48-670)

Active Caspase 3Cleaved PARPGAPHD

APOPTOSIS CELL SIGNALING KITSeveral proteins play key roles in the transduction of apop-

totic signals within a cell. The ability to monitor the abundance

of specific apoptotic pathway proteins, often simultaneously,

is necessary for understanding their role in cancer. The

MILLIPLEX MAP Apoptosis 3-Plex examines the expression of key

apoptosis proteins, as well as an analyte for protein normaliza-

tion.

OTHER SOLUBLE AND INTRACELLULAR CANCER BIOMARKERSIn addition to these panels developed specifically to study

cancer biomarkers, many of the analytes in our soluble and

intracellular panels have been used for the study of both

oncogenesis and the body’s response to tumor development.

These panels include:

• Cytokines and growth factors (IL-8, IL-10, HGF, etc.)

• Soluble cytokine receptors

• TGFβ• Adipokines (adiponectin, leptin, etc.)

• Bone metabolism

• Intracellular MAPmates (EGFR, Erb-B2, p53, JNK, etc.)

For a complete listing of analytes available, visit:

www.millipore.com/analytes.

CANCERYour Source for Advancing the Progress of Oncology Research

Our understanding of cancer is rooted in identifying

the phenotypic differences between cancer cells and

corresponding normal cells of the same lineage. Using data

obtained from clinical observations, basic cancer research

seeks to identify and characterize the mechanisms of

carcinogenesis—genetic and epigenetic changes that may

be involved in tumor development. In addition, in disciplines

such as molecular genetics, cell biology and biochemistry,

researchers explore the influences of heredity, diet and

environmental factors, using animal models and assays to

determine the function of genes and proteins that play a role

in tumor initiation, formation and invasion. These discoveries

have identified therapeutic candidates, which have then been

tested in clinical trials with the promise of benefiting cancer

patients. Millipore’s scientists share your determination to

conquer a disease that destroys the lives of so many. As your

steady partner in cancer research, we provide you with the

best soluble and intracellular multiplexing tools you need to do

your best work.

In recent years more research has focused on soluble cancer biomarkers. For example, understanding the role that both pro- and anti-inflammatory cytokines and chemokines play in the body’s immune response to cancer cells has shed light on the interactions between a tumor and its environment.

HUMAN CIRCULATING CANCER BIOMARKER PANEL 1(Cat. No. HCCBP1MAG-58K)

IL-6IL-8AFPCA125CA15-3 CA19-9CEACYFRA21-1FGF2/bFGFHCGβHE4HGFLeptin

MIF OsteopontinProlactinPSA (free)PSA (total) SCFSDF-1a + β/CXCL12sFas sFASLTGFaTNFaTRAILVEGF

NOTE: Catalogue numbers appearing in dark green font denote our magnetic bead format.

17

15

These graphs demonstrate the detection of these circulating cancer biomarkers in normal samples as well as lung and

prostate cancer panels.

Human Circulating Cancer Biomarker Magnetic Bead Panel (HCCBP1MAG-58K)

Con

cent

rati

on

4

3

2

1

0

200

150

100

50

Normal

CYF

RA

21

-1FG

F2 IL6

IL8

Prl

Lept

inCA

12

5sF

asO

PNsF

asl

TN

F-α

MIF

AFP

HG

FH

CG

-βH

E4CA

19

-9CA

15

-3VEG

FSCF

TR

AIL

TG

F-α

Toal

PSA

CEA

Detection of circulating cancer biomarkers in normal, lung and prostate cancer samples. Concentrations for TNFa, TGFa and TRAIL are in pg/mL.Concentrations for CA125, CA19-9 and CA15-3 ARE IN U/mL. All other analytes are in ng/mL.

Con

cent

rati

on

30

25

20

15

10

5

4

3

2

1

0

Lung Cancer

CYF

RA

21

-1FG

F2 IL6

IL8

Prl

Lept

inCA

12

5sF

asO

PNsF

asl

TN

F-α

MIF

AFP

HG

FH

CG

-βH

E4CA

19

-9CA

15

-3VEG

FSCF

TR

AIL

TG

F-α

Toal

PSA

CEA

Con

cent

rati

on

1500

10000

5000

0

Prostate Cancer

CYF

RA

21

-1FG

F2 IL6

IL8

Prl

Lept

inCA

12

5sF

asO

PNsF

asl

TN

F-α

MIF

AFP

HG

FH

CG

-βH

E4CA

19

-9CA

15

-3VEG

FSCF

TR

AIL

TG

F-α

Toal

PSA

CEA

CANCER

16

Cells respond to different stimuli in different ways using combinatorial,

reversible phosphorylation of signaling proteins. To understand cell

signaling pathways fully, it is not enough to determine if a protein is

phosphorylated or not—its function depends on:

• Which sites are phosphorylated?

• What fraction of the total protein population is phosphorylated?

Multiplexing, using the Luminex xMAP platform, provides faster answers

to these cell signaling questions compared to traditional Western blots,

mass spectrometry analyses and radioactive phosphorylation assays that

require large amounts of sample.

Multiplexed immunoassays advance the understanding of intracellular

communication by enabling the researcher to:

• Quantify multiple total and phosphorylated sites along a signaling

pathway, in a single sample

• Simultaneously quantify sites in multiple pathways

• Conduct all necessary measurements using a tiny sample volume

7-Plex T Cell Receptor Signaling Kit(Cat. No. 48-690)

CD3ε (pan Tyr)CREB (Ser133)Erk/MAPK 1/2 (Thr185/Tyr187)LAT (pan Tyr)Lck (pan Tyr)Syk (pan Tyr)ZAP-70 (pan Tyr)

8-Plex Multi-Pathway Signaling Kit (Cat. No. 48-680)

CREB (Ser133)Erk/MAPK 1/2 (Thr185/Tyr187)IkBa (Ser32)JNK (Thr183/Tyr185)p38 (Thr180/Tyr182)p70 S6 Kinase (Thr412)STAT3 (Ser727)STAT5A/B (Tyr694/699)

Cell Signaling is part of a complex system of communication that regulates cellular activity and pathways. Understanding the processes of cell signaling and the interaction of its pathways is essential in the process of unraveling and potentially eradicating metabolic disease, autoimmune disease and cancer.

CELL SIGNALINGYour Source for Moving Intracellular Research Forward

MILLIPLEX MAP CELL SIGNALING PORTFOLIOWith Millipore’s MILLIPLEX MAP Cell Signaling portfolio you choose the

product that best meets your needs: comprehensive panels in a one-

catalog-number, all-in-one kit format, an ever expanding portfolio of

MAPmates, or SignalProfiler, which features the expertise of our Lead

Discovery team. No matter what you choose, you receive the advantage of

speed and sensitivity, as well as ease-of-use, beginning with our simplified

ordering process and including our user-friendly protocols.

In recent years, more research has focused on soluble cancer biomark-ers. For example, understanding the role that both pro-and anti-inflammatory cytokines and chemo-kines play in the body’s immune response to cancer cells has shed light on the interactions between a tumor and its environment.

HUMAN MAPK/SAPK - 10 PLEX (Cat. No. 48-660)

Atf2 (Thr69/71)Erk/MAP Kinase 1/2 (Thr185/Tyr187)Hsp27 (Ser78)JNK (Thr183/Tyr185)c-Jun (Ser73)Mek1 (Ser222)Msk1 (Ser212)p38 (SAPK2A/B) (Thr180/Tyr182)p53 (Ser15)STAT1 (Tyr707)

HUMAN SRC FAMILY KINASE - 8 PLEX (Cat. No. 48-650)

Blk (pan Tyr)Fgr (pan Tyr)Fyn (pan Tyr)Hck (pan Tyr)Lck (pan Tyr)Lyn (pan Tyr)Src (pan Tyr)Yes (pan Tyr)

HUMAN STAT PANEL (Cat. No.48-610)

STAT1 (Tyr701)STAT2 (Tyr690)STAT3 (Tyr705)

STAT5A/B (Tyr694/Tyr699)STAT6 (Tyr641)

AKT/MTOR SIGNALING KIT (Cat. No. 48-611)

Akt (Ser473)GSK3a (Ser21)GSK3β (Ser9)IGF1R (Tyr1135/1136)IR (Tyr1162/1163)IRS1 (Ser312)

mTOR (Ser2448)p70 S6 Kinase (Thr412)PTEN (Ser380)RPS6 (Ser235/236)TSC2 (Ser939)

Cell Signaling Kits

Human Apoptosis Signaling Kits (Cat. No. 48-670)

Active Caspase 3 Cleaved PARPGAPHD

17

Because of the integral role of the Akt/mTOR pathway in many cellular processes, dysregulation of targets in multiple

pathways that signal through mTOR result in a variety of disease states, including chronic inflammatory disease and cancer.

The MILLIPLEX MAP 11-plex Akt/mTOR panel uses the traditional Luminex xMAP-based platform to detect simultaneously

changes in 11 pathway targets, further advancing both research and therapeutic strategies.

Human Akt/mTOR 11 Plex (48-611)

Effects of inhibition and stimulation of HepG2 cells on Akt/mTOR phosphorproteins. To demonstrate the power of the Akt/mTOR panel, HepG2 cells were treated with specific inhibitors such as wortmannin, which blocked phosphorylation of mTOR, TSC2, Akt, GSK3a, GSK3β, p70S6K and RPS6. This was followed by stimulation with 50 ng/mL IGF1 or 1 μM insulin for 15 minutes. Various phosphorylated biomarkers were simultaneously de-tected by the Luminex xMAP platform (represented as Average MFI) using Millipore’s Akt/mTOR panel.

2,000

p70S6

KIRS

GSK3α

IGF1R

GSK3β

AktPT

EN IRRPS

6TSC

2mTOR

0

MFI

4,000

6,000

HepG2 + InhibitorsHepG2 + Insulin

p70S6K

NT EGF(A431)

NT IGF(HEK293)

NT IGF(HEK293)

NT IGF(HEK293)

NT IGF(HEK293)

NT IGF(HEK293)

NT Serum(HEK293)

NT IGF(MCF-7)

NT IGF(MCF-7)

NT Ins(HepG2)

NT Ins(HepG2)

IRS1 GSKα IGF1R GSK3β Akt PTEN IR RPS6 TSC2 mTOR

Confirmatory immunoprecipitation/Western blots (IP/IB). To demonstrate the specificity of the Akt/mTOR assay, 50 μg of lysates (nontreated (NT) or treated with 100 ng/mL EGF, 50 ng/mL IGF, 10 μg/mL insulin (Ins), or serum re-addition) were mixed with capture antibodies to immunoprecipitate each respective protein. The immunoprecipitated proteins were separated on SDS-PAGE, transferred to nitrocellulose, and probed with biotin-labeled phospho-specific detection antibodies. The proteins were imaged using streptavidin-HRP and chemiluminescence.

CELL SIGNALING

18

Design Your Own Intracellular Signaling Panel with MILLIPLEX MAP MAPmatesIf you prefer to design a custom assay, Millipore has simplified

this process for you, making ordering the necessary kit compo-

nents easier so that you are assured you have everything you

need to do your best work.

Step 1: Plan your assayChoose the analytes you want to plex together and determine

their compatibility. Keep in mind that the following cannot be

plexed together:

o Total and phosphorylated MAPmate pairs

o Pan Tyr and phospho-specific MAPmates

o Phospho-MAPmates for a single target (Akt, STAT3, p53)

Using the following buffer table, analytes that use the same

assay buffer can be included in your custom assay. This enables

you to concentrate on proteins belonging to a specific signal

transduction pathway or to look at downstream targets that

have a role in multiple pathways.

Step 2: Order the MILLIPLEX maP Cell Signaling Buffer and Detection KitOrder the MILLIPLEX MAP Cell Signaling Buffer and Detection Kit

(Catalogue No. 48-602). With the exception of MAPmates, this

buffer kit contains everything you need to run your assay.

Step 3: Order the MAPmates you have selectedA MAPmate consists of:

• Capture Antibody (bead)

• Detection Antibody (biotin)

• Positive Control (stimulated cell lysate) for

phosphorylated analytes

Available MAPmates:

Cat. No. Control Sample Cat. No. Control Sample

47-205 HeLa: unstim 47-215 HEK293: LiCl

47-206 Jurkat: unstim 47-217 Daudi: IL-4

47-207 Jurkat: anisomycin 47-218 A549: camptothecin

47-208 Jurkat: H2O

247-219 3T3: anisomycin

47-210 A431: EGF 47-221 3T3: unstim

47-211 HeLa: HS/Ars 47-222 HeLa: HGF

47-212 HeLa: TNFa 47-224 Ramos: Pervanadate

47-213 NIH3T3: PDGF 47-225 HL-60: Pervanadate

47-214 Kasumi-3: SCF 47-226 HeLa: IFNa

Cell Lysates: Positive and Negative ControlsOur stimulated and unstimulated cell lysates are designed to be

used as controls with MILLIPLEX MAP Cell Signaling Kits and

MAPmates. These lysates are also recommended for use in

Western blotting and immunoprecipitation.

ANALYTE TOTAL PHOSPHORYLATED

Akt/PKB (assay buffer 1) 3 3 (Ser473)

Akt/PKB (assay buffer 1) 3 (Thr308)

Akt/PKB (assay buffer 2) 3 3 (Thr308)

Akt/PKB (assay buffer 2) 3 (Ser473)

ATF2 3 3 (Thr69/71)

BAD 3 3 (Ser112)

Bcl-2 3 3 (Ser70)

Active Caspase 3 3

CREB 3 3 (Ser133)

EGF-Receptor 3 3 (PanTyr)

Erb-B2/HER2 3 3 (PanTyr)

Erk/MAPK 1/2 3 3 (Thr185/Tyr187)

GSK3β 3 3 (Ser9)

H2A.X 3 (Ser139)

HIF-1a 3

HSP27 3 3 (Ser78)

IGF1R 3 3 (PanTyr)

IkBa 3 3 (Ser32)

IR 3 3 (Tyr1162/1163)

IRS1 3 3 (PanTyr)

JNK 3 3 (Thr183/Tyr185)

c-Jun 3 (Ser73)

c-Kit 3 3 (PanTyr)

Lck 3 3 (PanTyr)

c-Met/HGFR 3 3 (PanTyr)

MEK1 3 3 (Ser222)

MKK6 3 3 (Ser207/Thr211)

MSK1 3 3 (Ser212)

mTOR 3 3 (Ser2448)

p21 3

p38 (SAPK2A/B) 3 3 (Thr180/Tyr182)

p53 3 3 (Ser15)

p53 3 (Ser46)

p70 S6 Kinase 3 3 (Thr412)

Cleaved PARP 3

PDGF Receptor a 3 (PanTyr)

PDGF Receptor β 3 (PanTyr, mouse)

PRAS40 3 3 (Thr246)

PTEN 3 3 (Ser380)

Rsk1 3 (Ser380)

STAT1 3 3 (Tyr701)

STAT2 3 3 (Tyr690)

STAT3 3 3 (Tyr705)

STAT3 3 (Ser727)

STAT5A/B 3 (Tyr694/699)

STAT6 3 (Tyr641)

ZAP-70 3 (PanTyr)

GAPDH 3 (Human)

CELL SIGNALING

19

Successful Multiplexing with MILLIPLEX MAP

EpiQuant TechnologyTo overcome multiplexing challenges, in addition to our

MILLIPLEX MAP cell signaling kits and MAPmates, Millipore

proudly introduces our new MILLIPLEX MAP EpiQuant cell signal-

ing portfolio—a major advance in the design and implementa-

tion of quantitative immunoassays.

Richer Data from EpiQuant Sample ProcessingA unique feature of the EpiQuant sample processing protocol

is fragmentation or digestion of the protein sample. This pro-

cess has a number of benefits:

• Protein:protein interactions are disrupted

• A synthetic peptide can be used as a quantitative standard

• Specificity of the antibodies can be narrowed to a small

linear peptide sequence

• Cross-reactivity with similar peptide sequences in other

proteins can be predicted and avoided

• Residual enzymatic activity of the sample

(e.g., phosphatases and proteases) is quenched

Details of MILLIPLEX maP EpiQuant ProductsThe MILLIPLEX MAP EpiQuant platform unites two powerful

technologies: the EpiQuant technology which uses computa-

tional algorithms to guide the development of highly specific

antibodies and the Luminex xMAP platform that provides a

bead-based, high throughput, multiplex analysis. Answering

the challenges of multiplex cell signaling, MILLIPLEX MAP

EpiQuant assays enable you to:

• Design small- or large-plexed panels easily

• Customize your assay by selecting analytes you want to

study

• Obtain quantitative results at picomolar sensitivities

• Measure phosphorylation concentrations of multiple sites

on the same protein—all in one well

• Measure both total and phosphoprotein—all in the same

well

MILLIPLEX MAP EPIQUANT CELL SIGNALINGWhile multiplexed immunoassays using the Luminex xMAP

platform have advanced the understanding of intracellular

communication, multiplexing still presents some challenges.

There is a lack of phosphospecific antibodies that work well on

the Luminex platform due to:

• Requirement for native protein conformation instead of

recognition of denatured proteins on Western blots

• Complex buffer interactions with phosphoproteins

• Matrix effects as well as inter-antibody effects

when multiplexed

Resulting difficulties include:

• Lack of sufficient targets for pathway analysis

• Qualitative, not quantitative results

• The need for separate wells to analyze each phosphor-

ylated site on a specific analyte, as well as total

protein.

CELL SIGNALING

CELL SIGNALING

BIOLOGICAL VALIDATION FOR MILLIPLEX MAP EPIQUANT ANALYTESAll MILLIPLEX MAP EpiQuant analytes have been validated for both biological relevance and specificity.

MILLIPLEX MAP EpiQuant technology computational algorithms to identify unique, continuous linear sequences in proteins, generating anti-bodies against these specific sequences. Proteolytic digestion of your samples liberates these EpiQuant-bearing peptides, enabling you to simultaneously measure both total protein abundance and site-specific phosphorylation at the peptide level, using EpiQuant antibodies. These peptides are then used to generate standard curves, which results in quantitative measurements with picomolar sensitivity.

CBL (Y700)

CDCP1 (Y734)

CRK (Y221)

EGFR (Y1110)

EZRIN (Y477)

FAK (Y397)

FAK (Y861)

GHR (Y566)

HER2 (Y1023)

IL15Ra (Y227)

IRS1 (Y612)

ITK (Y180)

JAK3 (Y785)

JNK (Y185)

MET (Y1003)

p38 (Y181)

PDK1 (Y373)

PLAK (Y548)

PLCg1 (Y1253)

PLCg2 (Y1197)

RAF1 (Y340)

SCF38 (Y20)

SDCP3A (Y231)

SDCP3A (Y95)

SHC (Y349)

SPTAN1

TEC (Y206)

TFR (Y20)

WASP (Y291)

Con

cent

rati

on (pM

)

0

20

40

60

80

100

120

140

H 0

EGF

Non

2 2

Phosphotyrosine Activation in Unstimulated and Stimulated A431 Cells

CELL SIGNALING

20

CELL SIGNALING

PHOSPHOTYROSINE CELL SIGNALING PANEL 1 (Cat. No. MPEQ-100K)

Cbl (Tyr700/731/774)Crk (Tyr221/239CUB domain containing protein 1 (Tyr734)EGFR (Tyr1110/1125)Ezrin (Tyr477)FAK (Tyr397/407FAK (Tyr861)FRS2 (Tyr436)Fyb (Tyr97/651)Gab1 (Tyr285/307/317)Gab2 (Tyr266)Gab2 (Tyr584)Growth hormone receptor (Tyr566)

HER2 (Tyr1023)HGFR (Tyr1003)IL-15Ra chain (Tyr227)IRS1 (Tyr612)ITK (Tyr180)JAK3 (Tyr785)JNK 1/2/3 (Tyr185/185/223)LAT (Tyr255)p38/MAPKa (Tyr181)PDK1 (Tyr373/376)PI3K p110g (Tyr485)Plakoglobin/g-catenin (Tyr548)PLCg 1 (Tyr1253)PLCg 2 (Tyr1197/1217)

Raf-1 (Tyr340/341)Rap GEF1/GRF2 (Tyr504)SCF38 (Tyr20)SH2 domain containing protein 3A (Tyr231)SH2 domain containing protein 3A (Tyr95)Shc (Tyr349/350)SPTAN1 (Tyr1176)STAT3 (Tyr705)Transferrin Receptor (Tyr20)Tyrosine protein kinase Tec (Tyr206)VEGFR1 (Tyr1169)WASP (Tyr291)ZAP70 (Tyr126)

MILLIPLEX MAP EPIQUANT SAMPLE PREPARATION KIT (MPEQ-SP*)• A sample prep kit containing all the reagents you need to

prepare up to 5 mL of sample lysates, including:

Cell lysis

Protein denaturing

Protein fragmentation

• *Kit required to run MILLIPLEX map EpiQuant multiplex

assays

Now available, MILLIPLEX MAP EpiQuant panels with 40 phosphotyrosine analytes (see below). Future panels will include other sets

of total and phosphotyrosine-, phosphoserine- and phosphothreonine-modified analytes. Plus, pathway-specific assays enabling

you to explore at great depth pathways such as EGFR and p53.

MILLIPLEX MAP EPIQUANT PHOSPHOTYROSINE CELL SIGNALING PANEL 1 (MPEQ-100K)• All the components and reagents you need to

simultaneously detect multiple phosphotyrosine sites in

a small sample size

• The ability to choose analytes from different pathways

to design a custom kit

• Standards included to obtain quantitative picomolar-

range results

• Quality controls provided to qualify assay performance

• Analytically validated panels that yield consistent analyte

profiles irrespective of plex size

• Standardized standard curve ranges across analytes and

lots to ensure lot-to-lot consistency

• Panels that meet stringent manufacturing criteria to

ensure batch-to-batch reproducibility

CELL SIGNALING

21

Using GeneGo Inc. the above targets have been assigned to the following pathways:

Target Apo

ptos

is/S

urvi

val

Dev

elop

men

t Pa

thw

ays

Imm

une

Res

pons

e

Che

mot

axis

EPO

Act

ivat

ion

Cel

l Adh

esio

n

Cyt

oske

leto

n Rem

odel

ing

Reg

ulat

ion

of L

ipid

Met

abol

ism

G-P

rote

in S

igna

ling

EGFR

/ E

rbB D

evel

opm

ent

Sign

al T

rans

duct

ion

Tra

nscr

ipti

on

Tra

nsla

tion

Tra

nspo

rt

Cbl (Y700/731/774)

Crk (Y221/239)

CUB domain containing protein 1 (Y734)

EGFR (Y1110/1125)

Ezrin (Y477)

FAK (Y397/407)

FAK (Y861)

FRS2 (Y436)

Fyb (Y697/651)

Gab1 (Y285/307/317)

Gab2 (Y266)

Gab2 (Y584)

GHR (Y566)

HER2 (Y1023)

IL-15R (Y227)

IRS1 (Y612)

ITK (Y180)

JAK3 (Y785)

Jnk 123 (Y185/223)

LAT (Y255)

MET (Y1003)

p38 MAPKa (Y181)

PDK1 (Y373/376)

PI3K p100 (Y485)

Plakoglobin (g-catenin) (Y550)

PLCg1 (Y1253)

PLCg2 (Y1197/1217)

Raf-1 (Y340/341)

RapGEF1 (GRF2) (Y504)

SCF38 (Y20)

SH2 domain containing protein 3A (Y231)

SH2 domain containing protein 3A (Y95)

Shc (Y349/350)

SPTAN1 (Y1176)

STAT3 (Y705)

Tec (Y206)

TfR (Y20)

VEGFR1 (Y1169)

WASP (Y291)

ZAP70 (Y126)

CELL SIGNALING

22

MILLIPLEX map EpiQuant assays enable researchers to measure

phosphorylation levels of multiple sites, as well as total protein,

simultaneously with absolute quantitation. Based on the Luminex

xMAP platform, the EpiQuant technology uses antibodies against

defined protein sequences to capture peptides resulting from

protein linearization and digestion in a single cell lysate sample.

The fragments generated are similar to those generated using

mass spectroscopy. These same peptides are then used as stan-

dards to generate curves with picomolar sensitivity.

EXPLORING THE EGFR PATHWAY USING MILLIPLEX EPIQUANT EGFR SIGNALING MAGNETIC BEAD PANEL

EGFR SIGNALING PATHWAY PANEL (Cat. No. MPEQ110MAG-110K)

CSK (Tyr304)EGFR TotalEGFR/HER1 (Tyr1110/1125)EGFR/HER1 (Tyr1069/1092)EGFR/HER2 (Tyr1197/1199/1262/1276/1298/1307)ERK ½ (Try204/187)FAK (Tyr397/407)FAK (Tyr861)FRS2 (Tyr436)Gab1 (Tyr285/307/317)

Gab2 (Tyr266) Gab2 (Tyr584)Gab2 (Tyr614)PISKp 110δ (Try485)PKμ (Tyr502)PLCg1 (Tyr1253)PLCg2 (Tyr1192-1217)Shc (Tyr349/350)SPRY2 (Tyr55)STAT3 (Tyr705)TAFII68 Total (loading control)

Phosphorylated EGFR pathway proteins, as well as total EGFR, were simultaneously detected in A431 cells treated with 100 ng/mL EGF. Lysates were collected at 0, 5, 19, 20, 30, 45 and 60 minutes. Values are internally normalized utilizing the TAFII68 loading control.

3a 3b

3c 3d

Time (Min)

[Ana

lyte

] (p

M)

1,000

100

10

10 20 40 6010 30 50 70

ErbB3 (pY1197/1307) HER2 (pY1023)

EGFR (pY1110/1125) EGFR (pY1069/1092) EGFR total

[Ana

lyte

] (p

M)

Time (Min)

FAK (pY397/407) FAK (pY861) PKCm (pY502)

pl3Kp110d (pY485) PLCg1 (pY1253)

0.1

1

10

100

0 10 20 30 40 50 60 70

EpiQuant EGFR Panel: EGF Time Course

[Ana

lyte

] (p

M)

Time (Min)

FRS2 (pY436) Gab1 (pY285/307/317) Gab2 (pY266)

Gab2 (pY584) Gab2 (pY614)

0.1

1

10

100

0 10 20 30 40 50 60 70

[Ana

lyte

] (p

M)

Time (Min)

ERK1/2 (pY204/187) Shc (pY349/350) SPRY2 (pY55)

STAT3 (pY705) Csk (pY304)

0.01

0.1

1

10

100

1000

0 10 20 30 40 50 60 70

The complexity and number of protein targets involved in signaling events, as well as cellular responses, require tools that enable

multiplex analysis of samples. Millipore’s MILLIPLEX map EpiQuant technology allows for the analysis of a greater number of intracel-

lular analytes per well, as compared to even traditional Luminex intracellular assays. The magnetic bead format of these assays

provides additional convenience, with walk-away washing options, and increased consistency, with lower coefficients of variation.

The MILLIPLEX map EpiQuant EGFR pathway magnetic bead

panel enabled the detection of phosphorylation events for all

panel analytes in dose response and time course experiments. The

TAFII68 loading control values obtained were used to normalize the

amount of lysate for each well.

The figure below illustrates the results of the time course

experiments. The perpetuation of maximal signal is observed for

many targets (ERK1/2 and EGFR, for example) across the whole

of the time course, while for other targets (FRS2 and PKCμ, for

example) maximal signal levels are observed at the 5-minute time

point with a return to basal levels by 20 to 30 minutes. As would

be expected due to receptor internalization, it appears as though

total EGFR levels drop after stimulation, with a rebound to less

than resting levels at the one hour time point.

CELL SIGNALING

23

CELL SIGNALING ELISASMillipore is also your source for cell signaling ELISAs including:

• cAMP and cGMP High Throughput Screening (HTS) ELISAs

• Dual Detect CELISAs

CELL SIGNALING

Star ELISAs

ELISA - Other AssaysELISA - Cell Signaling / Intracellular

Description Cat. No.

AKT1 (total) 17-455

EGFR (total) 17-460

Erk 1/2 (total) 17-463

FAK (total) 17-479

GSK-3β (total) 17-471

IGF-1R (total) 17-481

Insulin Receptor (β subunit) (total) 17-483

IRS1 (total) 17-458

IkB (total) 17-485

JNK 1/2 (total) 17-465

MEK1 (total) 17-473

Met (total) 17-469

p38a (total) 17-487

p53 (total) 17-476

PRAS40 (total) 17-477

Src (total) 17-467

phospho-Akt (Ser473) 17-457

phospho-Akt (Thr308) 17-456

phospho-EGFR (Tyr1068) 17-462

phospho-EGFR (Tyr1173) 17-461

phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) 17-464

phospho-FAK (Tyr397) 17-480

phospho-GSK-3β (Ser9) 17-472

phospho-IGF-1R (Tyr1135/1136) 17-482

phospho-IR (β Subunit) (Tyr 1162/1163)

17-484

phospho-IRS1 (Ser307 mouse/Ser312 Human) 17-459

phospho-IkB (Ser32) 17-486

phospho-JNK 1/2 (Thr183/Tyr185) 17-466

phospho-MEK1 (Ser218/222) 17-474

phospho-MET (Tyr1230/Tyr1234/Tyr1235) 17-470

phospho-p38a (Thr180/Tyr182) 17-488

phospho-p53 (Ser15) 17-475

phospho-PRAS40 (Thr246) 17-478

phospho-Src (Tyr418) 17-468

DescriptionColorimetric

Cat. No.Chemiluminescent

Cat. No.

cAMP - 192 Assays 17-418

cAMP - 192 Assays 17-498

cAMP - 768 Assays 17-416

cAMP - 960 Assays 17-518

cAMP - 960 Assays 17-598

cAMP - 3840 Assays 17-516

cGMP - 192 Assays 17-419

cGMP - 768 Assays 17-417

EZ-Transcription Factor

HTS cAMP / cGMP

DescriptionColorimetric

Cat. No.Chemiluminescent

Cat. No.

AP-1 Family 70-550

ATF2 70-585 70-685

c-Fos 70-545 70-645

c-Jun 70-540 70-640

Creb (Ser133) 70-575 70-675

ErbB2 (Tyr1248) 17-490

FoxO3a 70-553

HDAC 17-374

Jun/Fos 70-546 70-646

NFkB Family 70-560 70-660

NFkB p50 70-515 70-615

NFkB p50/p65 70-510 70-610

NFkB p65 70-520 70-620

Oct-4 70-665

p53 70-525

PI3 Kinase Activity/Inhibitor 17-493

PhosphaFluor HTS OmniPhosphatase

17-448

Ras Activation 17-497

ssDNA Apoptosis APT225

STAT3 70-530 70-630

Universal TF 70-500 70-600

Universal TF 70-501 70-601

• G-Protein ELISA

• Signal Transduction Assay Reaction (STAR) ELISAs

• Transcription Factor (EZ-TFA) ELISAs

CELL SIGNALING

24

25

CELL SIGNALING

DescriptionColorimetric

Cat. No.Chemiluminescent

Cat. No.

cAMP - 192 Assays 17-418

cAMP - 192 Assays 17-498

cAMP - 768 Assays 17-416

cAMP - 960 Assays 17-518

cAMP - 960 Assays 17-598

cAMP - 3840 Assays 17-516

cGMP - 192 Assays 17-419

cGMP - 768 Assays 17-417

7000

6000

5000

4000

3000

2000

1000

0

Assa

y Bu

ffer

HeLa

: uns

tim

ulat

edHe

La: T

NFα

HeLa

: HGF

HeLa

: IFN

Jurk

at: u

nstim

ulat

ed

Jurk

at: a

niso

myc

inM

CF7:

IGF-

1NI

H3T3

: PDG

F

NIH3

T3: a

niso

myc

in

Jurk

at: H

2O 2

A431

:EGF

Cell and Stimulation

Experiment 1Experiment 2

MFI

Reproducibility of Erk-1/2 phosphorylation in multiple cell lines under various stimulation conditions.

Comparison of phosporylation states in HeLa cells upon HGF stimulation for 5 minutes.

Phos

pho

GSK3

α (S

er21)

Phos

pho

GSK3

β (S

er9)

Phos

pho

IGF1

R (Ty

r1135/T

yr1136)

Phos

pho

IRS (Ser

312)

Phos

pho

p70 S

6 K

inas

e (T

hr412)

Phos

pho

Akt

/PKB

(Ser

473)

Phos

pho

PTEN

(Ser

380)

Phos

pho

IR (Ty

r1162/1

163)

Phos

pho

RPS

6 (Ser

235/S

er236)

Phos

pho

TSC2 (Ser

939)

Phos

pho

mTO

R (Ser

2448)

Act

ive

Cas

pase

3To

tal G

APD

HCle

aved

PA

RP

(Asp

214)

Phos

pho

Bcl

-2 (Ser

70)

Phos

pho

p53 (Ser

15)

Phos

pho

p53 (Ser

46)

Phos

pho

Akt

/PKB

(Thr

308)

Phos

pho

Bad

(Ser

112)

Phos

pho

Rsk

1 (Ser

380)

Phos

pho

STA

T3 (Ty

r705)

Phos

pho

STA

T6 (Ty

r641)

Phos

pho

ERK 1

/2 (Thr

185/T

yr187)

Phos

pho

IkB

α (S

er32)

Phos

pho

MEK

1 (Ser

222)

Phos

pho

MSK1 (Ser

212)

Phos

pho

STA

T1 (Ty

r701)

Phos

pho

STA

T3 (Ser

727)

Phos

pho

STA

T5A

/B (Ty

r694/T

yr699)

Phos

pho

JNK/S

APK

1 (Thr

183/T

yr185)

Phos

pho

ATF2

(Thr

69/7

1)

Phop

ho c

-Jun

(Ser

73)

Phos

pho

p38/S

APK

(Thr

180/T

yr182)

Phos

pho

PRA

S40 (Thr

246)

Phos

pho

STA

T2 (Ty

r690)

Phos

pho

CR

EB (Ser

133)

Phos

pho

Lck

(Tyr

)Ph

osph

o PD

GF

Rec

epto

r α

(Tyr

)Ph

osph

o ZA

P-70 (Ty

r)Ph

osph

o c-

Kit

(Ty

r)Ph

osph

o c-

Met

/HG

FR (Ty

r)Ph

osph

o EG

F R

ec (Ty

r)Ph

osph

o Er

bB2/H

er2 (Ty

r)Ph

osph

o IG

F1R

(Ty

r)Ph

osph

o IR

S1 (Ty

r)Ph

osph

o Ly

n (T

yr)

Phos

pho

Src

(Ty

r)Ph

osph

o Ye

s (T

yr)

Phos

pho

Fgr

(Tyr

)Ph

osph

o Fy

n (T

yr)

Phos

pho

Blk

(Ty

r)Ph

osph

o H

ck (Ty

r)Ph

osph

o M

KK6 (Ser

207/T

hr211)

Phos

pho

Lat

(Tyr

)Ph

osph

o CD

3ε

(Tyr

)Ph

osph

o Syk

(Ty

r)

10500

8500

6500

45002500

2000

1500

1000

500

0

Analyte

Unstimulated

HGF

MFI

LEAD DISCOVERY SignalProfiler SERVICESProviding fresh insights to the cellular activity of

compoundsSignalProfiler service employs MILLIPLEX MAPmate cell sig-

naling 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 back-

grounds. In addition, early signaling events leading to cellular

toxicity can be studied to help assess potential compound

liabilities in early stages of drug development.

CELL SIGNALING

As a Luminex partner, Millipore is a distributor of Luminex instruments,

accessories and software. In fact, Millipore is the only partner to offer

Luminex xPONENT software integrated with a user-friendly, advanced data

analysis package, MILLIPLEX Analyst, to help you manage, analyze, and report

all your multiplex assays. We were there at the beginning to help validate

Luminex as the standard of multiplexing. And our ongoing dedication and

experience in this technology enable you to gain more information faster,

without compromising reliability.

GAIN MORE INFORMATION FASTER, WITHOUT COMPROMISING RELIABILITY• The Luminex platform and easy-to-use xPONENT and MILLIPLEX Analyst

software packages help you manage and track all of your multiplex

assays, enabling you to spend less time running your assays and more

time interpreting the significance of your data

• Benefit from the ability to quantitate up to 100 analytes in a single

sample simultaneously, saving you time, money and precious sample

• Choose from the broadest range of multiplex analytes for the Luminex

xMAP platform to meet your precise needs

• Advance your multiplexing research with integrated kits, instrumentation

and software

LUMINEX xMAP TECHNOLOGYYour Source for Simultaneous Multi-Analyte Detection

Millipore offers you the power of MILLIPLEX MAP multiplex immuno-detection kits and reagents developed by the industry’s original thought leaders for use with Luminex xMAP technology—one of the fastest-growing and most respected multiplex tech-nologies in drug discovery and development today.

The total system includes the Luminex 200 instrument, the Luminex XY plate handling platform and the Luminex SD sheath fluid delivery system and xPONENT 3.1 and MILLIPLEX Analyst software.

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INSTRUMENTATION & SOFTWARE

MAGPIX® SystemCombined with our MILLIPLEX mag magnetic bead-based

multi-analyte panels, analysis software, and technical support,

the new Luminex MAGPIX system provides a complete solution

for rapid, accurate biomarker quantitation in a variety of

sample matrices. This affordable system can perform up to

50 tests simultaneously in a single reaction volume, greatly

reducing sample input, reagents and labor while improving

productivity. The MAGPIX system features an innovative design

based on CCD imaging technology that allows for a more

compact, robust system. In addition it is easy to operate and

maintain with streamlined start-up and shutdown protocols

and minimal maintenance requirements.

Luminex 200™ – MILLIPLEX Analyzer The Luminex 200 System is a compact lab analysis system

that integrates: lasers, optics, fluidics, a controller, advanced

digital signal processing, XY Platform in a 17” x 20” footprint,

updated analysis software and a PC with monitor. Specifically

designed to enhance the use of xMAP technology, this system

enables simultaneous assays of up to 100 analytes per well of

a microtiter plate. Key features include: flexible data analysis

package, user friendly functions, flexible assay design

features, bar code reader for enhanced efficiency and multiple

formats for generating data reports.

For Research U

se Only.

Luminex FLEXMAP 3D™ SystemThe FLEXMAP 3D system combines differentially dyed

fluorescent microsphere sets with an innovative instrument

design that can accommodate measuring the concentrations

of up to 500 unique analytes within a single sample rapidly and

precisely. Features—such as automated probe height

adjustment, simplified routine maintenance operations and an

intuitive software interface—make FLEXMAP 3D a flexible and

easy-to-use multiplexing platform capable of performing a

variety of protein and nucleic acid applications.

NEW!

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INSTRUMENTATION & SOFTWARE

MILLIPLEX ANALYST SOFTWARE Data analysis software, MILLIPLEX Analyst, developed

exclusively for Millipore by VigeneTech, provides a whole

new level of data analysis, incorporating tried-and-true

algorithms to provide

the performance you

need, even when faced

with the most

challenging multiplexing

data sets. Also included

are report formats and

enhanced presentation

features that you’ve

requested to display

your information clearly

and accurately.

XPONENT® 3.1/4.0xPONENT 3.1/4.0 includes more than 65 new features

designed to benefit scientists and research laboratory

professionals by simplifying laboratory workflows and

increasing productivity. Although the graphical user

interface follows the typical assay workflow, it can be

customized to meet the needs of the specific laboratory.

Use of navigation wizards and automation of routine

operations (such as startup, shutdown, calibration and

performance verification) further enhance system usability

and result in increased walk-away time. xPONENT 3.1 is now

available on new Luminex 200 system or as an upgrade for

existing instruments. xPONENT 4.0 is available on the new

Luminex FLEXMAP 3D system.

For

Res

earc

h U

se O

nly.

HANDHELD MAGNETIC SEPARATOR BLOCK FOR 96 WELL FLAT BOTTOM PLATES OR 96 WELL CONICAL WELL PLATESThe Handheld Magnetic Separator Block offers a low cost

option for biomagnetic separation. Magnets are positioned

around the bottom

of the well to pull the beads to the

side of the well. The top magnetic

frame is white polycarbonate,

with a corrosion resistant

steel plate underneath, all

mounted to a polypropylene

base. An adjustable clip system

holds a wide variety of microplates

to the separator block, and an O-Ring

on the base plate facilitates gripping.

ELX405RM MICROPLATE WASHER FOR 96-WELL BIOMAGNETIC SEPARATIONIn collaboration with BioTek Instruments, the world leader

in microplate washing, Millipore introduces the newest

ELx405 enhancement—full plate biomagnetic separation

and washing for magnetic beads used in a growing number

of multiplex assays and bead-based ELISAs. The ELx405RM

combines superior microplate washing performance with

user-friendly features, making it an ideal washer for 96-well

microplate applications.

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MORE THAN THE BEST MULTIPLEXING PRODUCTS, WE’RE YOUR SOURCE FOR A COMPLETE WORKFLOW SOLUTIONBeyond products alone, we provide complete assay services all the way

through clinical trials. By obtaining all of the products and services you need

from a single source, you’ll have a continuity of knowledge that’s unachiev-

able any other way. Millipore’s BioPharma and BioMarker Services division is

the first to fundamentally redefine the concept of the CRO to help you

obtain better results, faster.

ARE YOU NEW TO MULTIPLEXING?Millipore can help. We offer several programs designed to help you quickly

familiarize yourself with the Luminex xMAP technology.

ADVANTAGES OF WORKING WITH MILLIPORE’S REGULA-TORY COMPLIANT LAB & BIOMARKER SERVICES TEAM:• Expertise in high quality sample analyses

• Fast, flexible reporting

• Guaranteed reproducibility and quality control

• Leadership in multiplex services using the Luminex xMAP Technology

• Largest portfolio available—choose from over 200 analytes

• Biomarker testing analysis for human, mouse, rat, primate

and canine samples

• GLP or Non-GLP services available

• Single or duplicate analysis

COMING IN 2011: THE RULES ARE CHANGING!Look to Millipore for state of the art, large research panels to support your

biomarker needs, in either singlet or duplicate analysis.

BIOPHARMA AND BIOMARKER SERVICESYour Source for Quality Services

What if you had a second lab to lend a hand with research and development projects? With more than 25 years of experience performing immunoassays on both human and non-human samples, Millipore’s BioPharma and Biomarker Services is here to support your laboratory projects and outsourcing needs. From RIAs, ELISAs and IRMAs (immunoradiometric assays) to multiplex assays using Luminex xMAP technology, our expert staff can work with you to determine the best assay format for your project. Whether you choose a Millipore assay or an assay developed by a third party, the flexibility of our BioPharma and Biomarker Services Teams means we’re able to meet your needs with comprehensive solutions and quality control.

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www.millipore.com/offices

Millipore, Advancing Life Science Together and MILLIPLEX are registered trade-marks of Millipore Corporation.ChemiKine, EpiQuant, MAPmates, SignalProfiler, ToxReporter and the M mark are trademarks of Millipore Corporation.Luminex, xPONENT, MAGPIX and xMAP are registered trademarks of Luminex Corporation.Luminex 200 and FLEXMAP 3D are trademarks of Luminex Corporation.FSC is a registered trademark of the Forest Stewardship Council AC.PB1096EN00 Rev. E Printed in U.S.A. 12/10 LS-SBU-10-03853© 2010 Millipore Corporation, Billerica, MA 01821 All rights reserved

TO PLACE AN ORDER OR RECEIVE TECHNICAL ASSISTANCE In the U.S. and Canada, call toll-free 1 800-Millipore (1-800-645-5476)

In Europe, please call Customer Service:

France: 0825.045.645

Spain: 901.516.645 Option 1

Germany: 01805.045.645

Italy: 848.845.645

English UK: 0870.900.46.45

For other countries across Europe and the world,

please visit www.millipore.com/offices.

For Technical Service, please visit www.millipore.com/techservice.