Oncology Product Selection Guide

8/6/2019 Oncology Product Selection Guide

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Data SheetProduct Selection Guide

Oncology Product Selection Guide

ONCOLOGY

TNF-

IL-8

AFP

Your Source for Comprehensive

Multiplex Panels for the Study

of Circulating and Intracellular

Cancer Biomarkers

CA15-3

MIF

HE4CEA

OPN

VEGF

SDF-1
http://www.millipore.com/bmia/flx4/bmia_cancer&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/?cid=BIOS-S-EPDF-1103-1107-DS


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WHATS INSIDE

Circulating CancerBiomarkers Panels and Kits

Other Circulating/SolubleBiomarker Panels and Kits

MILLIPLEXMAP CellSignaling Kits, MAPmatesand EpiQuant Panels

Calbiochem Top TenInhibitors

Luminex XMAP Technology


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Figure 1.Simultaneous Expression of Biomarkers in Serum Sample (Cat. No. 48-020)

Biomarker Expression Cancer Type

MIF Lung

Prolactin Breast, Prostate, and Ovarian

CA-125 Epithelial cancers

OPN Breast, Prostrate, Lung, Colon, Pancreas, Multiple myeloma, and Ovarian

Leptin Prostate and Breast

IGF-II Ovarian, Cervical, Colorectal, and Breast

Increased Expression Decreased Expression

Circulating Cancer Biomarkers Panels and Kits

The human cancer biomarker panel is a valuable research

tool for studying the biochemical mechanisms and signals

of disease, from early oncogenesis to metastasis. Using a

unique algorithm developed by Dr. Gil Mor of Yale University

in collaboration with Millipore, the expression prole of a

panel of six circulating biomarkers was used to distinguish

normal from ovarian cancer patient serum samples. This

panel includes both a ve-plex assay, which simultaneously

measures ve serum biomarkers; macrophage migration

inhibitory factor (MIF), prolactin, CA-125, leptin and

osteopontin (OPN), and a single-plex IGF-II assay.

The two graphs below demonstrate cancer biomarker

concentrations in normal versus ovarian cancer serum

samples. Cancer samples showed elevated levels of MIF,

prolactin, CA125 and osteopontin and reduced levels of

leptin and IGF-II, in agreement with ELISA results (data not

shown). The multiplex format delivers an efcient platform

for measuring multiple biomarkers simultaneously and

accurately, enabling you to save sample, time and money.

Human Cancer Biomarker Panel (48-020)

Differences in expression levels have been observed in many tumor types.
http://www.millipore.com/bmia/flx4/bmia_cancer&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-020&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-020&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-020&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer&cid=BIOS-S-EPDF-1103-1107-DS


5/28 3For more information, please visitwww.millipore.com/milliplex_cancer

Human Circulating Cancer Biomarker

Magnetic Bead Panel (HCCBP1MAG-58K)

ANALYTESBreast

Prostate

Lung

CRC

Pancreas

Leukemia

Lymphoma

Myeloma

Metastasis

Other

Cancer

(general)

Cancer

StemC

ells

AFP

Hepatocellular

Carcinoma,

Testicular orOvarian Tumor

CA15-3 Bone, Liver Ovary

CA19-9

CA125NSCL,

SCLOvary Endometria

CEA

CYFRA21-1

FGF2 (bFGF)NSCL,

SCL

Bladder, Melanoma

(Brain)

Bladder, GBM

(stem cells)Lung

HE4Ovary,

Endometrial

HCG Germ Cell Tumor,Testicular Tumor

HGF CML Prostate, BreastGetinib

ResistenceLung

IL-6B-CLL,

CML

Ovary, Drug

Resistance, H&N,

Gastric

Lung

IL-8 NSCL B-NHL Bladder

Bladder, Drug

Resistance, Ovary,

Esophagus

Hypoxia Lung

Leptin NSCL Uterine (EEC) CRC

MIF Ovary, Gastric

Osteopontin Breast Ovary

Prolactin OvarySCF Lung

SDF-1+/CXCL12 MALT

sFas Bladder

sFasLBladder, Esopha-

gus

TGF GBM, Sarcoma

TGF

AML,

MDS,

CML

Bladder, Ovary

Total/Free PSA

TRAIL Chemotherapeutic

VEGF Melanoma (Brain) Esophagus,Melanoma, Ovary Hypoxia Lung

In recent years more research has focused on soluble

cancer biomarkers. For example, understanding the role

that both pro- and anti-inammatory cytokines and

chemokines play in the bodys immune response to cancercells has shed light on the interactions between a tumor

and its environment.

Millipores MILLIPLEX MAP circulating cancer biomarker panel

consists of 26 circulating cancer biomarkers that can be

used to study a wide range of tumor types, examples of

which can be found in the table below. Some biomarkersare tumor type-specic, such as prostate-specic antigen

(PSA), while others, such as IL-8, have been detected in

many cancers.
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DS


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Concentration (pg/mL)

AFP

Leptin

Prolactin

CYFRA21-1

OPN

HE4

SDF1 +

MedianFluorescence

Intensity(MFI)100,000

10,000

1,000

100

10

1

010010 1,000 10,000 1,000,000100,000

44

ASSAY APPLICATIONS

This is an overnight incubation assay requiring 25 Lof 1:6 dilution plasma or serum, or 25 L cell culture

supernatant per well.

This kit may be used for the analysis of all or any

combination of the cancer biomarkers in serum, plasma

or tissue/cell lysate and culture supernatant with the

following exceptions:

Total PSA and Free PSA cannot be plexed together.

SDF1+ and CA19-9 cannot be plexed together.

SPECIFICITY

Cross-reactivity between the antibodies and any of theother analytes in this panel is non-detectable or negligible.

Assay Characteristics

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

MedianFlu

orescenceIntensity(MFI)100,000

10,000

1,000

100

10

1

01 10 100 1,000

Concentration (U/mL for CA15-3, 19-9, 125, mU/mL for HCG, pg/mL for IL-6, IL-8 and TNF)

CA 15-3

CA 19-9

IL-6

CA125

IL-8TNF

HCG

Median

FluorescenceIntensity(MFI)100,000

10,000

1,000

100

10

1

0

Concentration (pg/mL )

Total PSA

MIF

TRAIL

TGF-

Free PSA

VEGF

SFasL

SCF

CEA

FGF2

HGF

s Fas

101 100 1,000 100,00010,000

Figure 2.Circulating Cancer Biomarker Panel Magnetic Bead Curves
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DS


7/28 5For more information, please visit www.millipore.com/milliplex_cancer

AnalyteStandard

Curve Range(ng/mL)

Sensitivity+2SD

(ng/mL)

PrecisionAccuracy

(%)Intra-Assay(%CV)

Inter-Assay

(%CV)

AFP 0.3-250 83.4 6.0 6.7 86

CEA 0.02-20 5.2 11.3 4.6 74

CYFRA21-1 0.2-150 59.3 6.7 16.2 97

FGF2 (bFGF) 0.01-10 3.6 7.9 6.3 85

Free PSA 0.009-7 1.4 8.9 7.6 59

HE4 0.6-500 193.5 9.3 6.4 77

Total PSA 0.01-10 2.0 9.0 5.6 59

HGF 0.027-20 6.8 8.7 8.8 77

IL-6 0.0006-0.5 0.2 9.3 5.5 89

IL-8 0.001-1 0.3 6.8 5.5 51

Leptin 0.1-100 42.8 4.9 6.7 81

MIF 0.027-20 7.6 7.7 10.3 50

OPN 0.5-400 285.3 8.9 6.0 71

Prolactin 0.13-100 30.2 7.6 5.5 108

SCF 0.006-5 2.0 10.4 7.5 68

SDF1+ 0.1-100 33.9 12.2 15.4 35

sFas 0.034-25 8.4 6.5 7.1 69

sFasL 0.01-10 3.7 7.0 6.4 63

TGF 0.002-2 0.5 8.7 4.9 76

TNF 0.001-1 0.3 7.9 7.5 70

TRAIL 0.002-2 0.5 6.7 4.1 91

VEGF 0.01-10 6.4 12.0 10.2 74

Standard Curve RangeU/mL

Sensitivity +2SD(U/mL)

CA125 0.8-625 0.2 6.5 5.3 68

CA15-3 0.13-100 0.03 15.0 8.2 77

CA19-9 0.6-500 0.3 10.8 7.8 58

Standard Curve Range(mU/mL) Sensitivity +2SD(mU/mL)

HCG 0.09-67 29.5 7.5 5.3 69

ASSAY SPECIFICATIONS

Assay Characteristics

http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DS


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Cancer Biomarker Panels

Currently in Development

Future circulating cancer biomarker panels will includebroad tumor panels as well as those for the study of

angiogenesis and metastasis. The analytes included in

these panels will include:

Angiogenin

Antithrombin III

BMP-2

BMP-7

Extracellular matrix protein 1

Gelsolin

PLGF Thrombospondin-2

VEGF-C

Vitamin D binding protein precursor

Millipore assay development scientists are focused on

providing multiplex panels and kits that help you do your

best research.

The graphs at left demonstrate the detection of these

circulating cancer biomarkers in normal samples as well as

lung and prostate cancer panels.


Concentration

4

3

2

1

0

200

150

100

50

Normal

CYFRA21-1

FGF2IL6IL8Prl

Leptin

CA125

sFa

s

OPN

sFas

l

TNF-

MIF

AFP

HGF

HCG-

HE4

CA19-9

CA15-3

VEGF

SCF

TRAIL

TGF-

ToalPSA

CEA

Figure 3.Detection of circulating cancer biomarkers

in normal, lung and prostate cancer samples.

Concentrations for TNF, TGF and TRAIL are in pg/

mL.Concentrations for CA125, CA19-9 and CA15-3ARE IN U/mL. All other analytes are in ng/mL.

Concentration

30

25

20

15

10

5

4

3

2

1

0

Lung Cancer

CYFRA21-1

FGF2IL6IL8Prl

Leptin

CA125

sFas

OPN

sFasl

TNF-

MIF

AFP

HGF

HCG-

HE4

CA19-9

CA15-3

VEGF

SCF

TRAIL

TGF-

ToalPSA

CEA

Concentration

1500

10000

5000

0

Prostate Cancer

CYFRA21-1

FGF2

IL6

IL8

Prl

Leptin

CA125

sFas

OPN

sFasl

TNF-

MIF

AFP

HGF

HCG-

HE4

CA19-9

CA15-3

VEGF

SCF

TRAIL

TGF-

ToalPSA

CEA
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DS



Other Circulating /Soluble Biomarker Panels and Kits

In addition to the cancer biomarker panels, Millipores broad portfolio includes other multiplex assays that focus on cancer-

related analytes involved in inammation or metabolic disease. Measuring these biomarkers can help elucidate mechanisms

of oncogenesis and metastasis, accelerating your cancer research. These kits include:

AnalyteStandard

Curve Range(pg/mL)

Sensitivity+2SD

(pg/mL)

Precision

Accuracy(%)

Intra-Assay

(%CV)

Inter-Assay

(%CV)

Single Plex

TGF1 9.8 - 10,000 10 6 10 91

3-Plex

TGF1 9.8 - 10,000 12 5 13 83

TGF2 9.8 - 10,000 6 5 10 105

TGF3 9.8 - 10,000 6 5 9 46


TGF functions in angiogenesis, wound healing and

embryonic development. In addition, the TGF family plays a

critical role in immunity, heart disease, and cancer. Secreted

TGF cytokine exists in three isoforms: TGF1, TGF2 and

TGF3. This involvement in basic biological processes and

the opposing roles that the normal and mutated TGF

proteins play in tumor cell proliferation emphasize the

importance of this cytokine in a variety of research areas,

including cancer. The MILLIPLEX MAP multi-species TGF1

single plex and TGF 3-plex kits have been optimized for

performance and ease of use.

SPECIFICITY

TGF Single Plex:

This kit may be used to detect TGF1 in human, rat,

mouse, cynomolgus monkey, horse, rabbit, dog, cat,

guinea pig, pig and hamster. However, the amount of

species cross-reactivity has not been determined.

TGF3-plex:

TGF1 and TGF3 cross-reactivity with other analytes

in this panel is non-detectable or negligible. In this assay

TGF2 demonstrates some cross-reactivity with TGF1

(< 5%) and TGF3 (< 3%).

Human TGF1 may be used to detect TGF1 in rat,

mouse, cynomolgus monkey, horse, rabbit, dog, cat,

guinea pig, pig and hamster. However, the amount of

species cross-reactivity has not been determined.

Human TGF2 may be used to detect TGF2 in rat,

mouse, cynomolgus monkey, horse, rabbit, dog, guinea

pig, pig and hamster. However, the amount of species

cross-reactivity has not been determined.

TGF3 may not be detected in normal plasma and serum

samples. Due to the involvement of all three isotypes in

neonate development and lactation regulation, literature

suggests detectable levels in milk.

ASSAY RECOMMENDATIONS

Because platelet granules contain TGF1, which is

released upon platelet activation, collecting platelet-poorplasma samples is recommended.

TGF is secreted in a latent or inactive form; therefore,

serum, plasma and serum-containing culture samples

require acid treatment before testing to remove the

latency-associated peptide (LAP), releasing biologically

active mature TGF.

A 1:30 sample dilution is required for serum and plasma.

These are overnight assays requiring 25 L of sample.

Tumor Metastasis Panels

Multi-Species TGF1 Single Plex

(TGFB-64K-01)

Multi-Species TGF3-plex (TGFB-64K-03)
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-01&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-01&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-01&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-01&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-03&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-03&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-03&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-03&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-03&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/tgfb-64k-01&cid=BIOS-S-EPDF-1103-1107-DS


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The MMPs (matrix metalloproteinases), a family of zinc

proteases responsible for the breakdown of extracellular

matrix (ECM), play a key role in normal physiological

processes, such as embryonic development and tissue

morphogenesis, tissue and bone remodeling, wound healing

and angiogenesis.

The MILLIPLEX MAP human MMP panel 1 (MMP-3, -12, -13)

and MMP panel 2 (MMP-1, -2, -7, -9, -10) enable you to

explore modulation and function of MMP expression in

multiple therapeutic areas.

SPECIFICITY

No signicant cross-reactivity was observed among

analytes in each panel.


MMP Panel 1:

This is a 3.5-hour assay requiring 25 L neat human serum,

plasma, tissue/cell lysate and culture supernatant samples.

MMP Panel 2:

This is a 3.5-hour assay requiring 25 L 1:20 dilution

human serum, plasma, tissue/cell lystate and culture

supernatant samples.

AnalyteStandard

Curve Range(pg/mL)

Sensitivity+2SD

(pg/mL)

PrecisionAccuracy

(%)Intra-Assay

(%CV)

Inter-Assay

(%CV)

MMP Panel 1

MMP-3 49 50,000 32 5 13 MMP-3

MMP-12 98 100,000 105 7 11 MMP-12

MMP-13 9.8 10,000 41 6 20 115

MMP Panel 2

MMP-1 5 20,000 4 8 5 81

MMP-2 12 50,000 94 6 10 88

MMP-7 98 400,000 39 11 17 86

MMP-9 2.4 10,000 2 7 1 94

MMP-10 5 20,000 3 6 8 95


Tumor Metastasis PanelsHuman MMP Panel 1 (HMMP1-55K)

Human MMP Panel 2 (HMMP2-55K)
http://www.millipore.com/catalogue/item/hmmp1-55k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hmmp1-55k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hmmp2-55k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hmmp2-55k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hmmp2-55k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hmmp1-55k&cid=BIOS-S-EPDF-1103-1107-DS



Normal MMP function is precisely regulated by endogenous

protein inhibitors known as TIMPs (tissue inhibitors of

metalloproteinases), specically TIMP-1, TIMP-2, TIMP-3 and

TIMP-4. Disruption of the MMP/TIMP balance can result in

arthritis, cardiovascular disease, as well as tumor growth

and metastasis.

The MILLIPLEX MAP human TIMP panels enable you to explore

the modulation of TIMP expression and the function of the

MMP/TIMP balance in a variety of therapeutic areas. Human

TIMP panel 1 (TIMP-1, -2) is used for serum/plasma samples,

while human TIMP panel 2 (TIMP-1, -2, -3, -4) has been

designed for tissue/cell culture samples. Both panels are

customizable, enabling you to choose the analytes that best

t your needs.


MMP Panel 1:

This is an overnight or 2-hour assay requiring 25 L

1:50 dilution human serum or plasma samples. Overnightincubation is recommended.

MMP Panel 2:

This is an overnight or 2 -hour assay requiring 25 L urine,

tissue and cell culture samples. Overnight incubation is

recommended.

AnalyteStandard

Curve Range(pg/mL)

Sensitivity+2SD

(pg/mL)

Precision

Accuracy(%)

Intra-Assay

(%CV)

Inter-Assay

(%CV)

TIMP Panel 1 (serum/plasma)

TIMP-1 20 20,000 10 6 5 95

TIMP-2 49 50,000 19 7 9 93

TIMP Panel 2 (tissue/cell culture supernatants)

TIMP-1 20 20,000 10 6 5 96

TIMP-2 49 50,000 19 7 9 96

TIMP-3 98 100,000 101 9 6 85

TIMP-4 10 10,000 9 8 11 84


Tumor Metastasis PanelsHuman TIMP Panel 1 (HTIMP1-54K)

Human TIMP Panel 2 (HTIMP2-54K)
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/htimp1-54k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/htimp1-54k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/htimp2-54k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/htimp2-54k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/htimp2-54k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/htimp1-54k&cid=BIOS-S-EPDF-1103-1107-DS


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Insulin-like growth factor (IGF-1) is a polypeptide endocrine

hormone that is similar in structure to insulin. Stimulated

by growth hormone and produced primarily in the liver, it

binds to the IGF-1 receptor with much greater afnity thanto the insulin receptor, to which it binds with low afnity.

IGF-1 is involved in a wide range of therapeutic areas

including metabolic syndrome, growth deciency

and cancer.

Recognizing the severity of these disease states and the

therapeutic potential of studying IGF-1, Millipore provides

the MILLIPLEX MAP human and rat/mouse IGF-1 single

plex kits.


Human IGF-1 Single Plex:

This assay requires a sample extraction.

This assay requires 25 L per well of extracted serum

or plasma or tissue/cell culture samples. The tissue/cell

culture sample dilution may vary.

.

Rat/Mouse IGF-1 Single Plex:

This is an overnight incubation assay requiring 25 L per

well of 1:200 diluted serum or plasma samples or tissue /

cell culture samples.

Analyte

Standard

Curve Range

(pg/mL)

Sensitivity+2SD

(pg/mL)

Precision

Accuracy(%)

Intra-Assay

(%CV)

Inter-Assay

(%CV)

Human IGF-1 137 33,000 68 4 7 84

Rat/Mouse IGF-1 3.2 10,000 3.2 10 5 99


Metabolism and

Cancer PanelsHuman IGF-1 Single Plex (HIGF1-52K-01)

Rat/Mouse IGF-1 Single Plex (RMIGF187K)
http://www.millipore.com/catalogue/item/higf1-52k-01&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/higf1-52k-01&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/rmigf187k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/rmigf187k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/rmigf187k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/higf1-52k-01&cid=BIOS-S-EPDF-1103-1107-DS



ASSAY SPECIFICATIONS: Human IGFBP Panel

AnalyteStandard

Curve Range(ng/mL)

Sensitivity+2SD

(ng/mL)

Precision

Accuracy(%)

Intra-Assay

(%CV)

Inter-Assay

(%CV)

IGFBP-1 0.03 25 0.017


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Inammation contributes to a variety of disease states:

autoimmune, metabolic, cardiovascular, neurodegenerative

diseases and cancer. Emerging studies reveal a close

relationship between immune response and the

histopathology of cancer. These results t with the known

cooperation between tumor and non-tumor cells in the

process of tumor development. Many of these non-tumor

cells are immune cells, such as activated broblasts and

tumor-associated macrophages. Chronic inammation

mechanisms, such as the production of free radicals

usually associated with infection, have been shown to

foster tumorigenesis. In addition, pro-inammatory

cytokines such as TGFb, IL-6, IL-8 and TNF, as well as the

inammation NFB signaling pathway, play essential roles

in tumor biology.

With over 100 analytes available in a variety of species,

Millipores MILLIPLEX MAP cytokine/chemokine portfolio

provides all the tools needed to study the relationship

between inammation and cancer.

Response to chemotherapy is highly dependent on the

genetic prole of both the individual and the tumor.

MILLIPLEX MAP technology enables characterization of

multiple biomarkers associated with individual genetic

proles simultaneously. Preclinical studies of the vascular

targeting cancer drug candidate NSC 640488 demonstrated

two processes critical for anti-tumor effects: (1) tumor

vascular endothelial apoptosis and (2) production of

anti-vascular and pro-immunity cytokines1, 2. The same

research group used the MILLIPLEX MAP human cytokine

42-plex assay to identify a consistent pattern of cytokine/chemokine modulation in the peripheral blood lymphocytes

(PBL) of human donors that responded best to NSC 640488.

A small cohort of donors showed downregulation of IP-10,

MCP-1 and sCD40L and increased induction of TNF, MIP-

1, IL-6 and IL-8. This pattern of detection, shown in the

graphs below, may potentially correlate with a patients

responsiveness to NSC 640488: high responders (group A)

or low responders (group B).

1. Jassar AS et al. Activation of tumor-associated macrophages by the

vascular disrupting agent 5,6-dimethylxanthenone-4-acetic acid induces an

effective CD8+ T-cell-mediated antitumor immune response in murine models

of lung cancer and mesothelioma. Cancer Res. 2005 Dec 15;65(24):11752-61.

2. Ching LM et al. Induction of endothelial cell apoptosis by the antivascular

agent 5,6-Dimethylxanthenone-4-acetic acid. Br J Cancer. 2002 Jun

17;86(12):1937-42.

Inflammation and

Cancer Panels

Cytokine/Chemokine Panels

Human, Non-Human Primate,

Mouse, Rat, Canine

Inflammation and

Cancer Panels

Human Cytokine/Chemokine Magnetic

Bead Panel I (HCYTOMAG-60K)

Human Cytokine/Chemokine Panel I

(MPXHCYTO-60K)
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Figure 4.Cytokine Concentrations May Distinguish NSC 640488 Responders from Non-responders. The figure above

shows fold change in cytokine concentrations induced with NSC 640488 relative to untreated PBLs from different donors.

NC denotes less than a 1.5-fold change.


16/284

MILLIPLEX MAP Cell Signaling Kits,

MAPmates and EpiQuant Panels

The cells ability to respond to its environment is essential for normal development, cellular repair and immunity and is the

basis of homeostasis. Errors in these pathways, caused by mutations that may result in the constitutive expression or total

inhibition of protein transcription, are responsible for such diseases as diabetes, autoimmune disorders and cancer.

Using the Luminex xMAP platform, our MILLIPLEX MAP traditional bead-based cell signaling assays offer you the ability

to investigate simultaneously the expression of intracellular total or phosphorylated proteins involved in multi-pathway

signaling or focused in a specic pathway. Choose the format that best meets your needs: comprehensive panels in a

one-catalog-number, all-in-one kit format or an ever-expanding MAPmate portfolio.

Also using the Luminex xMAP platform, MILLIPLEX MAP EpiQuant technology differs from traditional multiplex cell signaling

assays because it can measure site-specic phosphorylation, enabling quantitative cell signaling analysis across pathways

and between multiple steps of a single pathway.

EpiQuant technology uses antibodies that have been generated against continuous linear peptide sequences in

phosphoproteins. Proteolytic digestion of your samples liberates these peptides, enabling you to measure simultaneouslyboth total protein abundance and site-specic phosphorylation at the peptide level. These peptides are then used to

generate standard curves, which results in quantitative measurements with picomolar sensitivity.

Figure 5.Effects of Small Molecule Activator on a

GPCR Cell-Line Lysate.

Initial cancer studies often require a survey of targets

from multiple cell signaling pathways. Millipores MILLIPLEX

MAP multi-pathway kit is an 8-plex assay that examines

phosphorylated proteins that span several signal

transduction pathways including MAPK, inammation and

apoptosis.

Figure 5 demonstrates the effects of a small molecule

activator on a GPCR cell line lysate. After incubation with

the activator, ChemiScreen GPCR cell line lysates were

analyzed using the human multi-pathway kit. The data

demonstrates dose-dependent phosphorylation of three

target proteins.

Analyte Phosphorylated

CREB (Ser133)

Erk/MAPK 1/2 (Thr185/Tyr187)

IB (Ser32)

JNK (Thr183/Tyr185)

p38 (Thr180/Tyr182)

p70 S6 Kinase (Thr412)

STAT3 (Ser727)

STAT5A/B (Tyr694/699)

MILLIPLEX MAP

Cell Signaling Kits Human Multi-Pathway 8 Plex (Cat.48-680)
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Often dysregulated during oncogenesis, proteins in this

pathway affect cellular homeostasis and response to the

extracellular environment, and are consequently targets

for drug development. The graphs below demonstrate theeffects of small molecules on phosphorylation of three cell

signaling proteins, underscoring the need to understand the

effect of drug candidates on key signaling pathways.


ATF2 (THR69/71)

ErK/MAPK 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)

Figure 6.Effects of small molecules inhibitors on

phosphorprotein in the MAP/SAPK pathway. The small

molecules U0126 (MEK1/2 inhibitor), U0124 (inactive form

of U0126), GW5074 (Raf-1 inhibitor) and SB203580 (p38

inhibitor) were used to study the effects of inhibition on the

phosphorylation of ERK, STAT1 and HSP27. ERK phosphoryla-

tion may be inhibited by a MEK1/2 inhibitor while there is no

effect on STAT1 (Figures 6a and 6b). STAT1 phosphorylation

may be inhibited by a Raf-1 inhibitor, while there appears to

be no effect on HSP27 phosphorylation (Figures 6b and 6c).

A p38 inhibitor seems to inhibit the phosphorylation of both

STAT1 and HSP27 (Figure 6c).

6A 6B

6C

MILLIPLEX MAP

Cell Signaling Kits Human MAP/SAPK 10 Plex (Cat.48-660)
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-660&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-660&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-660&cid=BIOS-S-EPDF-1103-1107-DS


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The Src family of proto-oncogenic kinases (SFK) is the

largest nonreceptor protein tyrosine kinase family

consisting of Src, Yes, Fyn, Fgr, Lck, Hck, Blk, and Lyn.

Expressed either ubiquitously or in hematopoieticimmune-competent cells, SFKs play crucial roles in

embryonic development and cell growth, regulating such

functions as proliferation, adhesion, differentiation, and

survival. Consequently, these signaling proteins are often

fundamentally involved in the progression and oncogenesis

of tumors such as non-small cell lung cancer (NSCLC),

squamous cell head and neck and pancreatic cancers.


Blk (pan Tyr)

Fgr (pan Tyr)Fyn (pan Tyr)

Hck (pan Tyr)

Lck (pan Tyr)

Lyn (pan Tyr)

Src (pan Tyr)

Yes (pan Tyr)

MILLIPLEX MAP

Cell Signaling Kits Human Src Family Kinase 8 Plex (48-650)

Figure 7a and 7b.

SFK phosphorprotein expression in

non-stimulated and stimulated cell lines.

To demonstrate the specificity of the MILLI-

PLEX MAP human Src family kinase 8 Plex, tumor

cell lines (Ramos, HL-60, HeLa and DLD-1) were

treated with pervanadate for 5 minutes. Cell ly-

sates prepared from untreated and pervanadate-

treated cells were incubated with a mixture of

all eight antibody-coated microbeads. Tyrosine

phosphorylation of SFKs was detected with a pan

antiphosphotyrosine antibody. The data showthat the multiplex assay readily detected tyrosine

phosphorylation: Blk, Fgr and Fyn phosphorylation

in Ramos cells; Lck, Lyn and Hck phosphorylation

in HL-60 cells, and Yes and Src phosphorylation in

HeLa and DLD-1 cells, respectively. Representative

multiplex assay data are shown in Figure 7a. The

confirmatory immunoprecipitation/Western blots

(IP/IB) shown in Figure 7b.

MedianFluorescenceIntensity

4,500

4,000

3,500

3,000

2,500

2,000

1,500

1,000

500

0Blk Fgr Fyn Lck Lyn Hck Yes Src

Pervanadate-treated cellsNon-stimulated cells

7a

BlkIP Antibody

Pervanadate

Fgr Lck LynRamos

+- +- +- +-

IP: Specific Capture Ab Blot: Phosphotyrosine Ab

Ramos HL-60 HeLaFynIP Antibody

Pervanadate

Hck Yes Src

+- +- +- +-

7b
http://www.millipore.com/catalogue/item/48-650&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-650&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-650&cid=BIOS-S-EPDF-1103-1107-DS



Mammalian Target of Rapamycin (mTOR), a serine/threonine

protein kinase, regulates production of signaling proteins

involved in key cellular processes, including:

RNA Polymerase I and III: protein synthesis/translation,cell metabolism

Cyclin D1: cell proliferation

p21: cell cycle progression following DNA damage,

apoptosis

HIF-1 and : angiogenesis and cell metabolism,

proliferation, motility, adhesion and survival

VEGF-C: lymphangiogenesis

Akt signaling is initiated by mitogenic stimuli resulting from

the binding of circulating growth factors to receptors such

as the insulin receptor (IR), the platelet-derived growth

factor receptor (PDGFR) and the epidermal growth factor

receptor (EGFR). In addition, Akt inhibits mTOR activity

either directly, by phosphorylating mTOR itself, or indirectly

through the tuberous sclerosis complex (TSC1/TSC2).

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 inammatory disease andcancer. 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.


Akt (Ser473)

GSK3 (Ser21)

GSK3 (Ser9)

IGF1R (Tyr1135/1136)

IR (Tyr1162/1163)

IRS1 (Ser312)

mTOR (Ser2448)

p70 S6 Kinase (Thr412)

PTEN (Ser380)

RPS6 (Ser235/236)

TSC2 (Ser939)

MILLIPLEX MAP

Cell Signaling Kits Human Akt/mTOR 11 Plex (48-611)

Figure 8.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,

GSK3, GSK3, p70S6K and RPS6.

This was followed by stimulation with

50 ng/mL IGF1 or 1 M insulin for

15 minutes. Various phosphorylated

biomarkers were simultaneously de-tected by the Luminex xMAP platform

(represented as Average MFI) using

Millipores Akt/mTOR panel.

2,000

p70S

6K IRS

GSK3

IGF1

R

GSK3

Ak

t

PTEN IR

RPS6

TSC2

mTO

R0

MFI

4,000

6,000

HepG2 + Inhibitors

HepG2 + Insulin
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-611&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-611&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-611&cid=BIOS-S-EPDF-1103-1107-DS


20/288

Analyte TOTAL Phosphorylated

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

Akt/PKB (assay buffer 1) (Thr308)

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

Akt/PKB (assay buffer 2) (Ser473)

ATF2 a (Thr69/71)

BAD a (Ser112)Bcl-2 a (Ser70)

Active Caspase a (Thr180/Tyr182)

CREB a (Ser133)

EGF-Receptor a (PanTyr)

Erb-B2/HER2 a (PanTyr)

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

H2A.X (Ser139)

HIF-1 a

HSP27 a (Ser78)

IGF1R a (PanTyr)

IB a (Ser32)

IR a (Tyr1162/1163)

IRS1 a (PanTyr)

JNK a (Thr183/Tyr185)

c-Jun (Ser73)

c-Kit a (PanTyr)

Lck a (PanTyr)

c-Met/HGFR a (PanTyr)

Analyte TOTAL Phosphorylated

MEK1 a (Ser222)

MKK6 a (Ser207/Thr211)

MSK1 a (Ser212)

mTOR a (Ser2448)

p21 a

p38 (SAPK2A/B)a

(Thr180/Tyr182)p53 a (Ser15)

p53 (Ser46)

p70 S6 Kinase a (Thr412)

Cleaved PARP a

PDGF Receptor (PanTyr)

PDGF Receptor (PanTyr, mouse)

PRAS40 a (Thr246)

PTEN a (Ser380)

Rsk1 (Ser380)

STAT1 a (Tyr701)

STAT2 a (Tyr690)

STAT3 a (Tyr705)

STAT3 (Ser727)

STAT5A/B (Tyr694/699)

STAT6 (Tyr641)

ZAP-70 (PanTyr)

GAPDH a(Human)

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 GSK IGF1R GSK3 Akt PTEN IR RPS6 TSC2 mTOR

In addition to the seven cell signaling kits proled within this brochure, Millipore provides you with the ability to design your

own kit with the buffer and detection kit (48-602) and your choice of MILLIPLEX MAPmates.

MILLIPLEX MAP

Cell Signaling Kits MILLIPLEX MAP Cell Signaling MAPmates

Figure 9.Confirmatory immunoprecipitation/Western blots (IP/IB). To demonstrate the specificity of the Akt/mTOR assay,

50 g of lysates (nontreated (NT) or treated with 100 ng/mL EGF, 50 ng/mL IGF, 10 g/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.



Analytes Phosphorylated

Cbl (Tyr700/731/774)

Crk (Tyr221/239)

CUB domain containing

protein 1 (Tyr734)PDK1 (Tyr373/376)

CUB domain containingprotein 1

(Tyr734)

EGFR (Tyr1110/1125)

Ezrin (Tyr477)

FAK (Tyr397/407)

FAK (Tyr861)

FRS2 (Tyr436)

Fyb (Tyr97/651)

Gab1 (Tyr285/307/317)

Gab2 (Tyr266)

Gab2(Tyr584)

Growth hormone receptor (Tyr566)

HER2 (Tyr1023)

HGFR (Tyr1003)

IL-15R chain (Tyr227)

IRS1 (Tyr612)

ITK (Tyr180)

JAK3 (Tyr785)

Protein Kinase Tec a(Tyr206)

ITK (Tyr180)

LAT (Tyr255)

Analytes Phosphorylated

JAK3 (Tyr785)

JNK 1/2/3 (Tyr185/185/223)

JNK 1/2/3 ZAP70 (Tyr185/185/223)

LAT (Tyr255)

p38/MAPKa (Tyr181)

PDK1 (Tyr373/376)

PI3K p110d (Tyr485)

Plakoglobin/g-catenin (Tyr548)

PLCg1 (Tyr1253)

PLCg2 (Tyr1197/1217)

Raf-1 (Tyr340/341)

Rap GEF1/GRF2 (Tyr504)

SCF38 (Tyr20)

SH2 domain containingprotein 3A

(Tyr231)

SH2 domain containingprotein 3A

(Tyr95)

Shc (Tyr349/350)

SPTAN1 (Tyr1176)

STAT3 (Tyr705)

Transferrin Receptor (Tyr20)

Tyrosine Protein Kinase Tec (Tyr206)

WASP (Tyr291)

ZAP70 (Tyr126)

Signaling events initiated by extracellular receptor

activation lead to the expression and release of cytokines

and chemokines. These events are central to cellular andtissue homeostasis, development and functional response.

The complexity and number of signaling proteins involved in

cellular responses require multiplex analysis of samples.

A549 lung carcinoma cells were cultured, then stimulated

with either EGF or TNF. For cytokine/chemokine analysis

tissue culture media was collected at various time points

over a 24-hour period, as well as unstimulated controls.

In addition, cell lysates were collected at time points over a

one-hour period. Prepared samples were assayed according

to the MILLIPLEX MAP human cytokine/chemokine-42-plex(MPXHCYTO60KPMX42) protocol and the MILLIPLEX MAP

EpiQuant phosphotyrosine cell signaling panel 1 (MPEQ-

100K) protocol, respectively. A total of 42 soluble analytes

and 105 intracellular phosphotyrosine proteins were

analyzed.

MILLIPLEX MAP EpiQuant

Cell Signaling Analysis

MILLIPLEX MAP EpiQuant

Phosphortyrosine Panel 1 (MPEQ-100K)

MILLIPLEX MAP EpiQuant Sample

Preparation Kit (MPEQ-SP)
http://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-100k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-100k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-100k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-100k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-100k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-sp&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-sp&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-sp&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-sp&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-sp&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-sp&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpeq-100k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DS


22/280

18

pM

FAK (PY397/407) EGFR (pY1110)

16

14

12

10

0

0 8 10 18 30 48 60

80

pM

60

40

20

0

0 8 10 18 30 48 60

EGFR (pY1110)40

pM

30

20

10

0

0 8 10 18 30 48 60

60 RET (pY10621090/1096)

pM

40

20

0

0 8 10 18 30 48 60

= TNF treated = EFG treated

time (minutes) time (minutes)

Figure 10.Site-specific phosphotyrosine concentrations over time (minutes) for several responsive phosphoproteins.

Of the 105 phosphotyrosine cell signaling targets analyzed, a response was observed in 38 target sites for TNF or EGF-treated

cells. The magnitude of cytokine/chemokine and phosphotyrosine response varied significantly between targets. For several phos-

phorylation sites, a decrease in phosphorylation was observed in response to TNF treatment.

pg/mL

IL-62000

1500

1000

500

0

1hr.

2hrs

.

4hrs

.

8hrs

.

16

hrs

.

24

hrs

.

pg/mL

IL-815000

10000

5000

0

1hr.

2hrs

.

4hrs

.

8hrs

.

16

hrs

.

24

hrs

.

pg/mL

FGF-280

40

20

0

1hr.

2hrs

.

4hrs

.

8hrs

.

16

hrs

.

24

hrs

.

pg/mL

IL-1015

10

5

0

1hr.

2hrs

.

4hrs

.

8hrs

.

16

hrs

.

24

hrs

.

= control = treated

Figure 11.Representative graphs of cytokine and chemokine expression. Analysis of TNF-stimulated A549 supernatant

revealed an increased expression in 21 of the 42 screened cytokine/chemokine analytes.



INHIBITORSELECT INHIBITOR LIBRARIES

AND SIGNALING PATHWAY PANELS

The Right Tools for Many ApplicationsFor decades, EMD Chemicals has manufactured high quality inhibitors, biochemicals, antibodies, proteins, and

kits that have been cited in thousands of peer-reviewed publications. To continue with our endeavor to offer

products that fast track your research, we have recently introduced the InhibitorSelect Libraries and Signaling

Pathway Panels that contain a collection of carefully selected, structurally diverse, and potent inhibitors in

convenient and cost-effective formats. InhibitorSelect products are suitable for a wide variety of applications

and are provided with comprehensive documentation.

FoldchangerelativetoD

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controlgrowthfactorback

ground

12

SU66

56

GSK-3Inh

ibitorX

III

Isogranula

timide

IC261

IKK-2

Inhib

itorIV

Indiru

binDeriva

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804

JNKInh

ibitorII

JNKInh

ibitor,Ne

gativ

eContro

l

JNKInh

ibitorV

JNKInh

ibitorIX

MK2a

Inhib

itor

JNKInh

ibitorV

III

K-25

2a,Noc

ardio

psiss

p.

Kenp

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MEKInh

ibitorI

MEKInh

ibitorII

MEK1/2Inh

ibitor

MNK1

Inhib

itor

NF-BAc

tivation

Inhib

itor

10

8

6

4

2

0

= No GFs = Sub EGF = Sub FGF2 = Max GFs

InhibitorSelect libraries are well characterized,

pharmacologically active, and structurally diverse

InSolution inhibitors in 96- or 384-well format.

These include:

96-well Library I (Cat. No. 539744)

Contains 80 inhibitors against mainly tyrosine, AGC and

atypical families of kinases in a volume of 50 L/well at a

concentration of 10 mM.

96-well Library II(Cat. No. 539745)

Contains 80 inhibitors against mainly CMGC and CaMK

families of kinases in a volume of 50 L/well at a

concentration of 10mM.

96-well Library III (Cat. No. 539746)

Contains 84 inhibitors against mainly CMGC, CaMK, AGC and

STE families of kinases, complementary to 96-well Library II,

in a volume of 50 L/well at a concentration of 10 mM.

384-well Library I (Cat. No. 539743)

Combines inhibitors from 96-well Library I and II for a total

of 160 inhibitors in one 384-well plate in a volume of 25

mL/well at a concentration of 10 mM.

InhibitorSelect Signaling Pathway panels consist of 12-16carefully hand-picked inhibitors in one convenient kit to

help you elucidate specic steps in a signaling cascade.

These include:

Signaling Pathway Panel PI3-K/Akt/mTOR

(Cat. No. 124031)

Signaling Pathway Panel EGFR (Cat. No. 324839)

Signaling Pathway Panel JAK/STAT (Cat. No. 420138)

Signaling Pathway Panel NFB (Cat. No. 481487)

Signaling Pathway Panel MAP Kinase

(Cat. No. 444189)

Signaling Pathway Panel Wnt (Cat. No. 681666)

For more information on InhibitorSelect products, pleasesee www.emdbiosciences.com/InhibitorSelect.com

Mouse Neural Stem Cell Viability

Figure 12.InhibitorSelect 96-Well Protein Kinase In-hibitor Libraries I & II. (160 inhibitors; Cat. No. 539744

and 539745) were screened for influence on proliferation

and survival of mouse neural stem cells (mNS) in an ATP

bioluminescence cell viability assay. mNS cells were plated

at 3,000 cells/well and viability assay performed after 2

days of incubation. Each compound was tested in each of

four growth factor backgrounds:

A. No GFs No Growth Factors (to identify survival/

proliferation factors)

B. Sub EGF Sub-optimal EGF (to identify inhibitors/

potentiators) 20pg/ml EGF

C. Sub FGF2 Sub-optimal FGF2 (to identify inhibitors/

potentiators) 500pg/ml FGF2

D. Max GFs Maximal EGF + FGF2 (to identify inhibitors/

potentiators) 20ng/ml EGF + 20ng/mL FGF2

Results for 20 inhibitors are presented as mean data (of

n=4 wells per condition) with error bars indicating standard

error of the mean (SEM). The presence of inhibitor K-252a,

Nocardiospsis sp. (Cat. No. 420297) alone in the culture

media resulted in a 10-fold mNS cell viability.

Data courtesy of Donna McLaren, Stem Cell Sciences,

Cambridge, UK
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BENEFITS OF LUMINEX TECHNOLOGY

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

signicance of your data

Benet 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 Technology

Millipore offers you the power of MILLIPLEX MAP multiplex immunodetection kits and reagents developed by the industrys

original thought leaders for use with Luminex xMAP technologyone of the fastest-growing and most respected multiplex

technologies in drug discovery and development today.

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 industry standard in multiplexing. And our ongoing dedication and experience in this technology

enable you to gain more information faster, without compromising reliability.
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LUMINEX INSTRUMENTS

Luminex MAGPIX

Millipore is proud to introduce Luminex MAGPIX instrumentation,

which combined with our MILLIPLEX mag magnetic bead-based

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

provides a complete solution for rapid, accurate biomarker

quantitation in a variety of sample matrices. This affordable system

can perform up to 50 test 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 enables for a more

compact, robust system. It is also 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, uidics, a controller, advanced digital

signal processing, and the XYP automation-capable platform in a 17x 20 footprint, updated analysis software and a PC with monitor.

Specically 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: exible data analysis

package, user friendly functions, exible assay design features,

bar code reader for enhanced efciency and multiple formats for

generating data reports.

Luminex FLEXMAP 3D

The FLEXMAP 3D system combines differentially dyed uorescent

microsphere sets with an innovative instrument design that can

accommodate measuring the concentrations of up to 500 uniqueanalytes within a single sample rapidly and precisely. Features

such as automated probe height adjustment, simplied routine

maintenance operations and an intuitive software interfacemake

FLEXMAP 3D a exible and easy-to-use multiplexing platform capable

of performing a variety of protein and nucleic acid applications.

SOFTWARE: Data Acquisition

xPONENT 3.1/4.0

xPONENT 3.1/4.0 includes more than 65 new features designed to benet scientists and research laboratory

professionals by simplifying laboratory workows and increasing productivity. Although the graphical user interface follows

the typical assay workow, it can be customized to meet the needs of the specic laboratory. Use of navigation wizards

and automation of routine operations (such as startup, shutdown, calibration and performance verication) 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.

SOFTWARE: Data Analysis

MILLIPLEX Analyst Software

The MILLIPLEX Analyst package 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 youve requested to display your information clearly

and accurately.
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26/284

Description Catalogue No.

Human Cancer Biomarker Panel 48-020

Human Circulating Cancer Biomarker

Magnetic Bead Panel

HCCBP1MAG-58K

Multi-Species TGF1 Single Plex TGFB-64K-01

Multi-Species TGF 3-Plex TGFB-64K-03

Human MMP Panel 1 HMMP1-55K

Human MMP Panel 2 HMMP2-55K

Human TIMP Panel 1 HTIMP1-54K

Human TIMP Panel 2 HTIMP2-54K

Human IGF-1 Single Plex HIGF1-52K-01

Rat/Mouse IGF-1 Single Plex RMIGF187K

Human IGFBP Panel HIGFBP-53K

Mouse IGFBP Magnetic Bead Panel MIGBPMAG-43K

Mouse IGFBP Panel MIGFBP-43K

Human Cytokine/Chemokine MagneticBead Panel I

HCYTOMAG-60K

Human Cytokine/Chemokine Panel I MPXHCYTO-60K

Human Multi-Pathway 8 Plex 48-680

Human MAP/SAPK 10 Plex 48-660

Human Src Family Kinase 8 Plex 48-650

Human Akt/mTOR 11 Plex 48-611

Cell Signaling Buffer and Detection Kit 48-602

EpiQuant Sample Preparation Kit MPEQ-SP

EpiQuant Human Phosphotyrosine Panel 1 MPEQ-100K

Featured Multiplex Products

ORDERING INFORMATIONMILLIPLEXMAP MAPmate Products96-well plate

MILLIPLEXMAP MAPmate Products96-well plate


Total Lck 46-617

Total JNK 46-618

Phospho c-Kit (Tyr) 46-619

Total c-Kit 46-620Total p21 46-621

Phospho c-Jun (Ser73) 46-622

Phospho STAT3 (Tyr705) 46-623

Phospho STAT3 (Ser727) 46-624

Total STAT3 46-625

Phospho IRS1 (Tyr) 46-627

Total IRS1 46-628

Phospho p70 S6 Kinase (Thr412) 46-629

Total p70 S6 Kinase 46-630

Phospho CREB (Ser133) 46-631

Total CREB 46-632Phospho STAT6 (Tyr641) 46-633

Total STAT2 46-634


Phospho STAT5A/B (Tyr694/

Tyr699)

46-641

Phospho IB (Ser32) 46-643

Total IB 46-644

Phospho Akt/PKB (Thr308),

Assay Buffer 1

46-645

Total IGF1R 46-646

Phospho IGF1R (Tyr) 46-647

Total ErbB2/Her2 46-648

Phospho ErbB2/Her2 (Tyr) 46-649

Total c-Met/HGFR 46-650

Phospho c-Met/HGFR (Tyr) 46-651

Total STAT1 46-654


Cleaved PARP (Asp214) 46-656

Total ATF2 46-657

Phospho ATF2 (Thr69/71) 46-658

Total Bcl-2 46-659

Phospho Bcl-2 (Ser 70) 46-660

Phospho ZAP-70 (Tyr) 46-661

Total p53 46-662

Phospho p53 (Ser15) 46-663

Phospho p53 (Ser46) 46-664

Total HIF-1 46-665

Total Human GAPDH 46-667

Total MEK1 46-669

Phospho MEK1 (Ser222) 46-670

Total MKK6 46-671

Phospho MKK6 (Ser207/Thr211) 46-672

Total MSK1 46-673

Phospho MSK1 (Ser212) 46-674


Phospho Akt/PKB (Ser473), Assay

Buffer 1

46-601

Phospho ERK 1/2 (Thr185/Tyr187) 46-602

Phospho EGF Rec (Tyr) 46-603

Active Caspase 3 46-604

Total Akt/PKB, Assay Buffer 1 46-605

Total EGF Receptor 46-606

Phospho Hsp27 (Ser78) 46-607

Total Hsp27 46-608

Total ERK 1/2 46-609

Phospho p38/SAPK

(Thr180/Tyr182)

46-610

Phospho Rsk1 (Ser380) 46-611

Total p38 46-612

Phospho JNK/SAPK1 (Thr183/

Tyr185)

46-613

Phospho PDGF Receptor (Tyr) 46-614

Phospho PDGF Receptor (Tyr) 46-615

Phospho Lck (Tyr) 46-616
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