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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 -
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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 -
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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
Human Circulating Cancer Biomarker Magnetic Bead Panel (HCCBP1MAG-58K)
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.
Human Circulating Cancer Biomarker Magnetic Bead Panel (HCCBP1MAG-58K)
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 -
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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
ASSAY SPECIFICATIONS
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.
ASSAY RECOMMENDATIONS
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
ASSAY SPECIFICATIONS
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 -
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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.
ASSAY RECOMMENDATIONS
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
ASSAY SPECIFICATIONS
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.
ASSAY RECOMMENDATIONS
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
ASSAY SPECIFICATIONS
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 -
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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)
http://www.millipore.com/catalogue/item/hcytomag-60k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hcytomag-60k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hcytomag-60k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpxhcyt0-60k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpxhcyt0-60k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/mpxhcyt0-60k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hcytomag-60k&cid=BIOS-S-EPDF-1103-1107-DS -
8/6/2019 Oncology Product Selection Guide
15/28 13For more information, please visit www.millipore.com/milliplex_cancer
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.
-
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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)
http://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-680&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-680&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/48-680&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_cancer#tab1=2&cid=BIOS-S-EPDF-1103-1107-DS -
8/6/2019 Oncology Product Selection Guide
17/28 15For more information, please visitwww.millipore.com/milliplex_cancer
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.
Analyte Phosphorylated
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.
Analyte Phosphorylated
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 -
8/6/2019 Oncology Product Selection Guide
19/28 17For more information, please visitwww.millipore.com/milliplex_cancer
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.
Analyte Phosphorylated
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 -
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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.
-
8/6/2019 Oncology Product Selection Guide
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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 -
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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.
-
8/6/2019 Oncology Product Selection Guide
23/28 2For more information, please visit www.millipore.com/milliplex_cancer
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.
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MSO
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ground
12
SU66
56
GSK-3Inh
ibitorX
III
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timide
IC261
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Inhib
itorIV
Indiru
binDeriva
tiveE
804
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ibitorII
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ibitor,Ne
gativ
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ibitorV
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ibitorIX
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itor
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ibitorV
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ibitorI
MEKInh
ibitorII
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ibitor
MNK1
Inhib
itor
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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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8/6/2019 Oncology Product Selection Guide
25/28 23For more information, please visitwww.millipore.com/milliplex_cancer
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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tp://www.millipore.com/bmia/flx4/bmia_software&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_software&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_software&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_software&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_software&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/catalogue/item/hccbp1mag-58k&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_software&cid=BIOS-S-EPDF-1103-1107-DShttp://www.millipore.com/bmia/flx4/bmia_instruments&tab2=2&tab2=2#tab2=2&cid=BIOS-S-EPDF-1103-1107-DS 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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
Description Catalogue No.
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 STAT2 (Tyr690) 46-635
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
Phospho STAT1 (Tyr701) 46-655
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
Description Catalogue No.
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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