Post on 10-Mar-2015
description
Data SheetProduct Selection Guide
MILLIPLEX® MAP and Cell Signaling PathwaysYour Source for Luminex® xMAP® Technology
IRS-1
Src PI3K Ras
STAT5 STAT2
STAT6 FAK
FynJAK2
JAK1
ZAP70
LckLAT
STAT1
GSK3 Akt
mTOR
Rsk
Raf
Mek1/2
Erk1/2
STAT1 STAT3 Rsk CREB ATF1 Histone 3
Msk1
p70 56k
ProteinSynthesis
Transcription
Stress
β-Catenin
BAD
Ras MKK3 IKK
IκBα
PRAK
Hsp27
NFκB
p38p38 JNK Bcl2
cdc25c
cdc2
Aurora 2
PARP
ATF2 Jun Smad2/3
MKK3/6MKK4/7
Smad2/3
FRHD
MYPT1
Apoptosis
BAD
Caspase 8
Caspase 3
Syk
Death
FASLTNFαR
TGFβRCD79β
CD79α
TGFβ1,2,3
slg
CD3ζCD3ε
Cytokines TCR
Growth
IGF-RIns-R
c-Metc-Kit
PDGF-RαPDGF-RβEGF-R
Brown = Available from Millipore
www.millipore.com/milliplex_csp
MILLIPLEX MAP andCell Signaling PathwaysDetermining the complex interactions between cell signaling pathways is essential for understanding how the cell communicates with and responds to its microenvironment to maintain homeostasis.
Determining how the elements of signal transduction interact both
within and between pathways is essential to understanding normal
and disease processes. Using the Luminex xMAP platform, MILLIPLEX
MAP Cell Signaling offers you the ability to investigate simultaneously
the expression of intracellular total or phosphorylated proteins
involved in multi-pathway signaling or focused in a specific pathway.
You choose what best meets your needs: comprehensive panels in
a one-catalog-number, all-in-one kit format or an ever-expanding
MAPmate™ portfolio. No matter what you choose, you receive the
advantage of speed and sensitivity, as well as ease of use, beginning
with our simplified ordering process and including our user-friendly
protocols.
Available Cell Signaling Portfolio
•MILLIPLEXMAPCellSignalingKits
•MILLIPLEXMAPMAPmateProducts
•MILLIPLEXMAPBufferandDetectionKit
•CellLysates
•SpeciesCross-reactivity
•ComparisonData
AnexampleofthepowerofourMILLIPLEXMAPCellSignalingPortfoliocomparedtoWesternblotsis
demonstratedinthegraphsbelow.JurkatcellsweretreatedwithC305anti-TCRantibodyoranisotype-
matchedcontrolantibodyat2µg/mLovera10-minuteperiod.TheMILLIPLEXMAPHumanTCellReceptorKit
wasusedtomeasuretheeffectsoftheantibodyversuscontrolonthephosphorylationofpanelanalytesover
time.TheaverageMFIandstandarddeviationof4replicatewellsareshown.
Sincebandintensitiescorrelatewithfoldincreases,theWesternpresentedinthegraphsvalidatethe
multiplexdata.ThisfiguredemonstratestheadvantageofusingMILLIPLEXMAPCellSignalingMAPmatesand
kits:
•ResultsareeasiertoquantitatethanWesterns.
•Multipletargetscanbemeasuredsimultaneously.
The Effects of an Anti-TCR Antibody on Phosphorylation in Jurkat Cells using the Human T Cell Receptor Kit (Catalogue No. 48-690)
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CD3ζ CD3ε
1 2 5 10
6789
10
2.52.01.51.0
00.5
3.03.54.04.55.0
10864
00
2Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
1214161820
10
5
00
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LCK ZAP70
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
15
20
25
2.52.01.51.0
00
0.5Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LAT Erk
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
2.52.01.51.0
00.5
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CREB
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
6789
10
5432
00
1Ave
rage
MFI
(1
00
0x)
Minutes of Stimulation
C305 Isotype Control
CD3ζ CD3ε
1 2 5 10
6789
10
2.52.01.51.0
00.5
3.03.54.04.55.0
10864
00
2Ave
rage
MFI
(1
00
0x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
1214161820
10
5
00
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LCK ZAP70
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
15
20
25
2.52.01.51.0
00
0.5Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LAT Erk
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
2.52.01.51.0
00.5
3.03.54.04.55.0
0
Ave
rage
MFI
(1
000x)
Minutes of Stimulation
C305 Isotype Control
CREB
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
6789
10
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CD3ζ CD3ε
1 2 5 10
6789
10
2.52.01.51.0
00.5
3.03.54.04.55.0
10864
00
2Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
1214161820
10
5
00
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LCK ZAP70
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
15
20
25
2.52.01.51.0
00
0.5Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LAT Erk
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
2.52.01.51.0
00.5
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CREB
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
6789
10
5432
00
1Ave
rage
MFI
(1
00
0x)
Minutes of Stimulation
C305 Isotype Control
CD3ζ CD3ε
1 2 5 10
6789
10
2.52.01.51.0
00.5
3.03.54.04.55.0
10864
00
2Ave
rage
MFI
(1
00
0x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
1214161820
10
5
00
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LCK ZAP70
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
15
20
25
2.52.01.51.0
00
0.5Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LAT Erk
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
2.52.01.51.0
00.5
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CREB
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
6789
10
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CD3ζ CD3ε
1 2 5 10
6789
10
2.52.01.51.0
00.5
3.03.54.04.55.0
10864
00
2Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
1214161820
10
5
00
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LCK ZAP70
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
15
20
25
2.52.01.51.0
00
0.5Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LAT Erk
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
2.52.01.51.0
00.5
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CREB
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
6789
10
5432
00
1Ave
rage
MFI
(1
00
0x)
Minutes of Stimulation
C305 Isotype Control
CD3ζ CD3ε
1 2 5 10
6789
10
2.52.01.51.0
00.5
3.03.54.04.55.0
10864
00
2Ave
rage
MFI
(1
00
0x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
1214161820
10
5
00
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LCK ZAP70
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
15
20
25
2.52.01.51.0
00
0.5Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
LAT Erk
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
2.52.01.51.0
00.5
3.03.54.04.55.0
0
Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
CREB
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
5432
00
1Ave
rage
MFI
(1000x)
Minutes of Stimulation
C305 Isotype Control
C305 anti-TCRβAntibody
Isotype ControlAntibody
1 2 5 10
1 minNT 2 min 5 min 10 min
6789
10
Cell signaling, often called signal transduction, is a complex system of
communication that enables cells to perceive, communicate with and respond to
their microenvironment. Some communication requires direct cell-to-cell contact.
Other effects are induced when a receptor that has been activated by a ligand
interacts directly with other proteins inside the cell. Still other cellular changes are
the result of more complex signal transduction pathways—kinase receptors that,
when bound to a ligand, become phosphorylated. In this case, the phosphorylation
can generate a binding site for a different secondary protein, inducing a protein-
to-protein interaction that enables signal transduction. A terminal signaling event,
such as a transcription factor, is phosphorylated, resulting in the activation that
ultimately affects, for example, cell cycle progression. These multi-component cell
signaling pathways are so complex that often there is feedback, signal amplification,
and interactions between more than one signal and among several signaling
pathways.
The cell’s 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.
MILLIPLEX MAP Cell Signaling Kits and MAPmate Panels
AVAILABLE ANALYTES/THERAPEUTIC AREAS* Oncology Inflammation
Metabolic Syndrome Autoimmune Stem Cells Apoptosis CNS
Akt/PKB • • • • •
ATF2 • • • • • • •
BAD • • • •
Bcl-2 • • •
Caspase3 • • •
c-Jun • • • • • •
c-Kit • • • • •
c-Met/HGFR • • • • •
c-Met/HGFR • • • •
CREB • • • • • •
EGFR • • • •
Erb-B2/HER2 • • • •
ERK1/2 • • • • • • •
GSK3b • • • • •
HistoneH2A.X • • •
Hsp27 • • • • • •
IGF-1R • • • •
IkBa • • • • • • •
IR • • • • •
IRS1 • • • • •
JNK • • • • • • •
Lck • • • • •
MEK1 • • • • • • •
MKK6 • • • • • • •
MSK1 • • • • • • •
mTOR • • • • •
p38/SAPK • • • • •
p53 • • • •
p70S6Kinase • • • • •
PARP • • •
PDGFRa • • • • • •
PDGFRb • •
PRAS40 • • • • •
PTEN(Ser380) • • • • •
Rsk1 • • • •
STAT1 • • • • • •
STAT2 • • • • • •
STAT3 • • • • • •
STAT5A/B • • • • • •
STAT6 • • • • • •
ZAP-70 • • • •
*BasedoncitationfrequencyinPubMed.
MILLIPLEX MAP Cell Signaling Kits
AnalytesfeaturedinMillipore’sMILLIPLEXMAPCellSignalingKitshavebeenanalyticallyvalidatedtogether.Eachkitincludes
allthereagentsnecessarytorunanassayon3525-µLsamplesinduplicate—allinonekit,withonecataloguenumber:
• Beads(captureantibodies)
• Biotin(detectionantibodies)
• Lysisbuffer
• Assaybuffer(s)
• Stimulatedandunstimulatedcelllysates
(positiveandnegativecontrols)
• Streptavidin-Phycoerythrin
• Opaquemulti-screenplateandcover
• Mixingvials
• User-friendlyprotocol
• Environmentally-friendlypackaging
Multi-Pathway – 8-Plex (Catalogue No. 48-680)MILLIPLEXmapMulti-PathwayKitisan8-plexassaythat
examineseightphosphorylatedproteinsthatspan
severalsignaltransductionpathwaysincludingMAPK,
inflammationandapoptosis.
Analyte Phosphorylated
CREB (Ser133)
Erk/MAPK1/2 (Thr185/Tyr187)
IkBa (Ser32)
JNK (Thr183/Tyr185)
Analyte Phosphorylated
p38 (Thr180/Tyr182)
p70S6Kinase(Thr412)
STAT3 (Ser727)
STAT5A/B (Tyr694/699)
Human T Cell Receptor – 7-Plex (Catalogue No. 48-690)MILLIPLEXmapTCellReceptorKitisa7-plexassaythat
examinessevenphosphorylatedproteinsinvolvedin
immuneresponse,tumordevelopmentandinflammation.
Analyte Phosphorylated
CD3e (panTyr)
CREB (Ser133)
Erk/MAPK1/2 (Thr185/Tyr187)
LAT (panTyr)
Analyte Phosphorylated
Lck (panTyr)
Syk (panTyr)
ZAP-70 (panTyr)
Human Apoptosis – 3-Plex (Catalogue No. 48-670)Apoptosisplaysafundamentalroleinmanynormalbiologicalprocesses,includingdevelopment,homeostasisandDNArepair.
MILLIPLEXmapHumanApoptosisKitiscurrentlya3-plexassaythatexaminesactivecaspase-3andcleavedPARP,proteins
involvedinimmuneresponse,tumordevelopmentandinflammation.GAPDHisprovidedasacontrolforproteinloading.
AdditionalMAPmateproductsarealsoavailableforthestudyofapoptosis.
ActiveCaspase3 GAPDHActiveCleavedPARP
Analytes
Human MAP Kinase – 10-Plex (Catalogue No. 48-660)ThecomplexMAPKinasepathwayconnectsintracellular
responsestothebindingofgrowthfactorsviacell
surfacereceptors.TheMILLIPLEXmapMAPKinaseKitisa
10-plexassaythatexaminestenphosphorylatedproteins
involvedinnormalmitosisandtumordevelopment.
Analyte Phosphorylated
ATF2 (Thr69/71)
ERK/MAP (Thr185/Tyr187)Kinase1/2
HSP27 (Ser78)
JNK (Thr183/Tyr185)
cJun (Ser73)
Analyte Phosphorylated
MEK1 (Ser221)
Msk1 (Ser212)
p38 (Thr180/Tyr182)
p53 (Ser15)
STAT1 (Tyr707)
Human Src Family Kinase Family (SFK) – 8-Plex (Catalogue No. 48-650)TheSrcfamilyofproto-oncogenickinases(SFK)isthelargestnonreceptorproteintyrosinekinasefamilyconsistingof:
Src,Yes,Fyn,Fgr,Lck,Hck,Blk,andLyn.Expressedeitherubiquitouslyorinhematopoieticimmune-competentcells,
membersoftheSFKregulatesuchfunctionsasproliferation,adhesion,differentiation,andsurvivalthatplaycrucialroles
inembryonicdevelopmentandcellgrowth.Consequently,thesesignalingproteinsareoftenfundamentallyinvolvedinthe
progressionandoncogenesisoftumorssuchasnon-smallcelllungcancer(NSCLC),squamouscellheadandneckand
pancreaticcancers.
Human Akt/mTOR – 11-Plex (Catalogue No. 48-611)TheAkt/mTOR(mammaliantargetofrapamycin)pathway
playsacentralroleinanumberofcellularprocesses
includingtranslation,cellcycle,cellularenergyhomeostasis
andapoptosis.Keyserineandthreoninekinases,Aktand
mTORareactivatedviagrowthfactorsignaling,promoting
increasedproteintranslationandcellgrowth.Growthfactors,suchasinsulinandIGF,promotephosphorylationoftheIR,
IGF1RandIRS1,aswellasthedownstreamtargetsAkt,GSK3a,GSK3b,p70S6K,andRPS6.Constitutivephosphorylationis
observedwithPTEN,TSC2,andmTOR.ComparedtoWesternblotting,resultsfromtheAkt/mTORsignalingpanelweremore
quantitativeandsensitiveandsavedtimeandresourceswhenusedtoanalyzephosphorylatedproteinbiomarkersincellular
lysates.
Blk
Fgr
Lck
Lyn
Src
Yes
Fyn
Hck
Analytes (all pan Tyr)
2,000
p70S6
KIRS
GSK3α
IGF1R
GSK3β
AktPT
EN IRRPS
6TSC
2mTOR
0
MFI
4,000
6,000
HepG2 + InhibitorsHepG2 + Insulin
To demonstrate the power of the Akt/mTOR panel, HepG2 cells were treated with specific inhibitors such as wortmannin, which blocked phosphorylation of mTOR, TSC2, Akt, GSK3a, GSK3b, p70S6K and RPS6. This was followed by stimulation with 50 ng/mL IGF1 or 1 μM insulin for 15 minutes. Various phosphorylated biomarkers were simultaneously detected by the Luminex® xMAP® platform (represented as Average MFI) using Millipore’s Akt/mTOR panel.
Analyte Phosphorylated
Akt (Ser473)
GSK3a (Ser21)
GSK3b (Ser9)
IGF1R (Tyr1135/1136)
IR (Tyr1162/1163)
IRS1 (Ser312)
Analyte Phosphorylated
mTOR (Ser2448)
p70S6Kinase(Thr412)
PTEN (Ser380)
RPS6 (Ser235/236)
TCS2 (Ser939)
MILLIPLEX MAP MAPmate Portfolio: Building Custom Intracellular Assays
Ifyouprefertodesignacustomassay,Milliporehassimplifiedthisprocessforyou,makingorderingthenecessarykit
componentseasiersothatyouareassuredyouhaveeverythingyouneedtodoyourbestwork.
Plan your assay
Choosetheanalytesyouwanttoplextogetheranddeterminetheircompatibility.Keepinmindthatthe
followingcannotbeplexedtogether:
oTotalandphosphorylatedMAPmatepairs
oPanTyrandphospho-specificMAPmates
oPhospho-MAPmatesforasingletarget(Akt,STAT3,p53)
Usingthefollowingbuffertable,analytesthatusethesameassaybuffercanbeincludedinyourcustom
assay.Thisenablesyoutoconcentrateonproteinsbelongingtoaspecificsignaltransductionpathwayor
tolookatdownstreamtargetsthathavearoleinmultiplepathways.
STEP 1.
Order the MILLIPLEX MAP Cell Signaling Buffer and Detection Kit (Catalogue No. 48-602)
WiththeexceptionofMAPmates,thisbufferkitcontainseverythingyouneedtorunyourassay.
Thekitincludes:
• MILLIPLEXMAPLysisBuffer
• MILLIPLEXMAPAssayBuffer1
• MILLIPLEXMAPAssayBuffer2
• MILLIPLEXMAPAmplificationBuffer
• Streptavidin-Phycoerythrin
• Negative/positivecontrol(Phosphorylated/Total)
• Opaque96-wellFilterPlateandcover
• Emptymixingvials
(BufferanddetectionkitisrequiredforMAPmateanalysisofsamples.)
STEP 2.
Order the MAPmate products you have selected
AMAPmateconsistsof:
• CaptureAntibody(bead)
• DetectionAntibody(biotin)
• PositiveControl(stimulatedcelllysate)forphosphorylatedanalytes
Thepositivecontrolcanbeusedtoqualifyassayperformanceor,withserialdilutions,
todevelopastandardcurve.
STEP 3.
ORDERING IS AS EASY AS 1-2-3. TheMILLIPLEXMAPbufferkitandMAPmateproductswillprovideyouwithallthecomponentsand
reagentsyouneedtosimultaneouslydetectthemultipleanalytesyouhavechosentostudy.
l Recommended • Acceptable 6 NotRecommended
Total MAPmate Portfolio
Catalogue No. Description Bead RegionLysis Buffer / Assay Buffer
1 2
46-605 TotalAkt/PKB,AssayBuffer1 38 l 6
46-675 TotalAkt/PKB,AssayBuffer2 62 6 l
46-657 TotalATF2 39 l l
46-695 TotalBad 59 l l
46-659 TotalBcl-2 61 l l
46-620 Totalc-Kit 8 l l
46-650 Totalc-Met/HGFR 46 l l
46-632 TotalCREB 37 l l
46-606 TotalEGFReceptor 34 l l
46-648 TotalErbB2/Her2 29 l l
46-609 TotalERK1/2 9 6 l
46-665 TotalHIF-1a 69 l l
46-608 TotalHsp27 50 • l
46-646 TotalIGF1R 48 l l
46-644 TotalIkBa 33 • l
46-687 TotalIR 65 l l
46-628 TotalIRS1 36 l l
46-618 TotalJNK 16 6 l
46-617 TotalLck 20 • l
46-669 TotalMEK1 45 l l
46-671 TotalMKK6 66 6 l
46-673 TotalMSK1 75 6 l
46-685 TotalmTOR 79 l l
46-621 Totalp21 28 l •46-612 Totalp38 54 6 l
46-662 Totalp53 53 6 l
46-630 Totalp70S6Kinase 19 l l
46-680 TotalPRAS40 55 l 6
46-678 TotalPTEN 63 • 6
46-654 TotalSTAT1 24 • l
46-634 TotalSTAT2 67 l l
46-625 TotalSTAT3 14 l •46-667 TotalHumanGAPDH 15 6 l
Forthemostupdatedversionofthistable,visitwww.millipore.com/milliplex_csp.Thenclickthe“MAPmateBufferTable”link.
Phosphorylated MAPmate Portfolio
Catalogue No. Description Bead RegionLysis Buffer / Assay Buffer
1 2
46-604 ActiveCaspase3 6 l l
46-656 CleavedPARP(Asp214) 60 l l
46-601 PhosphoAkt/PKB(Ser473),AssayBuffer1 38 l 6
46-645 PhosphoAkt/PKB(Thr308),AssayBuffer1 38 l 6
46-677 PhosphoAkt/PKB(Ser473),AssayBuffer2 62 6 l
46-676 PhosphoAkt/PKB(Thr308),AssayBuffer2 62 6 l
46-658 PhosphoATF2(Thr69/71) 39 l l
46-694 PhosphoBad(Ser112) 59 l •
46-660 PhosphoBcl-2(Ser70) 31 l l
46-622 Phosphoc-Jun(Ser73) 52 l •
46-619 Phosphoc-Kit(Tyr) 8 l l
46-651 Phosphoc-Met/HGFR(Tyr) 46 l l
46-631 PhosphoCREB(Ser133) 37 l l
46-603 PhosphoEGFRec(Tyr) 34 l l
46-649 PhosphoErbB2/Her2(Tyr) 29 l •
46-602 PhosphoERK1/2(Thr185/Tyr187) 9 6 l
46-692 PhosphoHistoneH2A.X(Ser139) 43 l •
46-607 PhosphoHsp27(Ser78) 50 l l
46-647 PhosphoIGF1R(Tyr) 48 l l
46-688 PhosphoIR(Tyr1162/1163) 65 l l
46-643 PhosphoIkBa(Ser32) 33 • l
46-627 PhosphoIRS1(Tyr) 36 l l
46-613 PhosphoJNK/SAPK1(Thr183/Tyr185) 16 6 l
46-616 PhosphoLck(Tyr) 20 • l
46-670 PhosphoMEK1(Ser222) 45 • l
46-672 PhosphoMKK6(Ser207/Thr211) 66 • l
46-674 PhosphoMSK1(Ser212) 75 6 l
46-686 PhosphomTOR(SER2448) 79 l l
46-610 Phosphop38/SAPK(Thr180/Tyr182) 54 6 l
46-663 Phosphop53(Ser15) 53 • l
46-664 Phosphop53(Ser46) 53 • l
46-629 Phosphop70S6Kinase(Thr412) 19 6 l
46-614 PhosphoPDGFReceptora(Tyr) 21 • l
46-615 PhosphoPDGFReceptorb(Tyr) 32 • l
46-681 PhosphoPRAS40(Thr246) 55 l 6
46-679 PhosphoPTEN(Ser380) 63 • l
46-611 PhosphoRsk1(Ser380) 42 l •
46-655 PhosphoSTAT1(Tyr701) 24 • l
46-635 PhosphoSTAT2(Tyr690) 67 l l
46-624 PhosphoSTAT3(Ser727) 14 l •
46-623 PhosphoSTAT3(Tyr705) 14 l •
46-641 PhosphoSTAT5A/B(Tyr694/Tyr699) 35 • l
46-633 PhosphoSTAT6(Tyr641) 10 l l
46-661 PhosphoZAP-70(Tyr) 51 • l
Forthemostupdatedversionofthistable,visitwww.millipore.com/milliplex_csp.Thenclickthe“MAPmateBufferTable”link.
l Recommended • Acceptable 6 NotRecommended
MILLIPLEX MAP GAPDHUnique to Millipore
TheGlyceraldehyde3-PhosphateDehydrogenase(GAPDH)geneisconstitutivelyand
stablyexpressedinalmostalltissuesandcells,andisconsideredtobea
“housekeeping”gene.
MilliporehasdevelopedahumanGAPDHMAPmatetoaidyouinyourcellular
experiments,usingitasaloadingcontrolandforproteinnormalizationinyour
multiplexassays.
6,000
05 2.5
Concentration (µg/mL)
Ave
MFI
10 0.00
8,000
4,000
2,000
HeLa
HEK293
A431
A549
Kasumi
MCF7
GAPDH Human Lysate Titrationusing Assay Buffer 2 Total
GAPDH
Jurkat HeLa
Cell Lysates: Positive and Negative ControlsOurstimulatedandunstimulatedcelllysatesweredesignedtobeusedascontrols
withMILLIPLEXMAPCellSignalingKitsandMAPmateproducts.Theselysatesarealso
recommendedforuseinWesternblottingandimmunoprecipitation.
Available Lysates Available Lysates
Catalogue No. Control Sample Catalogue No. Control Sample
47-205 HeLa:unstim 47-216 MCF-7:IGF
47-206 Jurkat:unstim 47-217 Daudi:IL-4
47-207 Jurkat:anisomycin 47-218 A549:camptothecin
47-208 Jurkat:H2O
247-219 3T3:anisomycin
47-210 A431:EGF 47-221 3T3:unstim
47-211 HeLa:HS/Ars 47-222 HeLa:HGF
47-212 HeLa:TNFa 47-224 Ramos:Pervanadate
47-213 NIH3T3:PDGF 47-225 HL-60:Pervanadate
47-214 Kasumi-3:SCF 47-226 HeLa:IFNa
47-215 HEK293:LiCl
GAPDHcellularconcentrationscanvarysignificantlydependingonthecellline.Aninitialsampletitrationsimilartothatshownintherepresentativedataaboveisrecommendedforanypreviouslyuntestedcellline.
3,000
00.03 0.06 0.13 0.25 0.50
µg of Cell Lysate per Well
MFI
0.01 0.02 1.00
4,0003,500
2,500
1,500
4,500
2,000
1,000500
Jurkat: unstimulated
Jurkat: anisomycin
MILLIPLEX MAP Detection of Cleaved PARP in Jurkat Cells
Western BlotCleaved PARP
– +anisomycin
IP/WesternCleaved PARP
– +anisomycin
ThegraphsbelowshowrepresentativedataobtainedwhenusingMillipore’scell
lysatesineachofthefollowingassays.Ineachcase,stimulatedandunstimulated
lysateswereresuspendedinultrapurewater.
• MILLIPLEX MAP: Aserialdilutionofarehydratedlysatewasanalyzedusingan
appropriateMAPmate.TheMedianFluorescenceIntensity(MFI)wasmeasuredwith
theLuminexInstrument.
• Western Blot: AliquotsoflysatewereanalyzedusingtheappropriateMAPmate
biotin-labeleddetectionantibodyandimagedusingstreptavidin-HRPand
chemiluminescence.
• Immunoprecipitation/Western: Aliquotsoflysatesweremixedwiththe
appropriateMAPmatecaptureantibodybeadstoimmunoprecipitatetheprotein.
TheimmunoprecipitatedproteinswereseparatedonSDS-PAGE,transferredto
nitrocellulose,andprobedwiththesameMAPmatebiotin-labeleddetection
antibody.Theproteinswereimagedusingstreptavidin-HRPandchemiluminescence.
3,000
00.03 0.06 0.13 0.25 0.50
µg of Cell Lysate per Well
MFI
0.01 0.02 1.00
4,0003,500
2,500
1,500
4,500
2,000
1,000500
Jurkat: unstimulated
Jurkat: anisomycin
MILLIPLEX MAP Detection of Cleaved PARP in Jurkat Cells
Western BlotCleaved PARP
– +anisomycin
IP/WesternCleaved PARP
– +anisomycin
Cell Lysates: Positive and Negative Controls (continued)
Total MAPmate Portfolio
Catalogue No. DescriptionSpecies Reactivity
Human Mouse Rat
46-605 TotalAkt/PKB,buffer1 l l l
46-675 TotalAkt/PKB,buffer2 l l l
46-657 TotalATF2 l l
46-695 TotalBad l l
46-659 TotalBcl-2 l
46-620 Totalc-Kit l
46-632 TotalCREB l l l
46-606 TotalEGFReceptor l
46-648 TotalErbB2/Her2 l
46-609 TotalErk1/2 l l l
46-665 TotalHIF-1a l l l
46-608 TotalHsp27 l
46-646 TotalIGF1R l
46-644 TotalIkBa l
46-628 TotalIRS1 l l l
46-618 TotalJNK l l
46-617 TotalLck l l
46-669 TotalMEK1 l l
46-650 Totalc-Met/HGFR l
46-671 TotalMKK6 l l l
46-673 TotalMSK1 l
46-621 Totalp21 l
46-612 Totalp38 l l l
46-662 Totalp53 l
46-630 Totalp70S6Kinase l l l
46-654 TotalSTAT1 l l
46-634 TotalSTAT2 l
46-625 TotalSTAT3 l l l
46-667 TotalHumanGAPDH l
46-687 TotalIR l l l
46-685 TotalmTOR l l l
46-678 TotalPTEN l l l
46-680 TotalPRAS40 l
Forthemostupdatedversionofthistable,visitwww.millipore.com/milliplex_csp.
Thenclickthe“SpeciesCross-ReactivityTable”link.
MILLIPLEX MAP Intracellular Assays: Species Cross-Reactivity
Whilemostofthecaptureanddetectionantibodiesareanti-human,manyare
cross-reactivewithotherspecies.Inmostcasestheseanalyteswereidentifiedby
examiningaminoacidsequencetodeterminehomology.Thetablebelowillustrates
speciescross-reactivityforeachMAPmateproduct.
Phosphorylated MAPmate Portfolio
Catalogue No. DescriptionSpecies Reactivity
Human Mouse Rat
46-601 PhosphoAkt/PKB(Ser473),buffer1 l l l
46-677 PhosphoAkt/PKB(Ser473),buffer2 l l l
46-645 PhosphoAkt/PKB(Thr308),buffer1 l l l
46-676 PhosphoAkt/PKB(Thr308),buffer2 l l l
46-658 PhosphoATF2(Thr69/71) l l
46-694 PhosphoBad l l
46-660 PhosphoBcl-2(Ser70) l
46-604 ActiveCaspase3 l l
46-631 PhosphoCREB(Ser133) l l l
46-603 PhosphoEGFR(Tyr) l
46-649 PhosphoErbB2/Her2(Tyr) l
46-602 PhosphoErk1/2(Thr185/Tyr187) l l l
46-692 PhosphoHistoneH2A.X l l
46-607 PhosphoHsp27(Ser78) l
46-647 PhosphoIGF1R(Tyr) l
46-643 PhosphoIkBa(Ser32) l
46-688 PhosphoIR(Tyr1162/1163) l l
46-627 PhosphoIRS1(Tyr) l l l
46-613 PhosphoJNK/SAPK1(Thr183/Tyr185) l l
46-622 Phosphoc-Jun(Ser73) l l l
46-619 Phosphoc-Kit(Tyr) l
46-616 PhosphoLck(Tyr) l l l
46-670 PhosphoMEK1(Ser222) l l l
46-651 Phosphoc-Met/HGFR(Tyr) l
46-672 PhosphoMKK6(Ser207/Thr211) l l
46-674 PhosphoMSK1(Ser212) l
46-686 PhosphomTOR(Ser2448) l l l
46-610 Phosphop38/SAPK(Thr180/Tyr182) l l l
46-663 Phosphop53(Ser15) l
46-664 Phosphop53(Ser46) l
46-629 Phosphop70S6Kinase(Thr412) l l l
46-656 CleavedPARP(Asp214) l
46-614 PhosphoPDGFReceptora l l
46-615 PhosphoPDGFReceptorb l
46-681 PhosphoPRAS40(Thr246) l
46-679 PhosphoPTEN(Ser380) l l l
46-611 PhosphoRsk1(Ser380) l l l
46-655 PhosphoSTAT1(Tyr701) l l
46-635 PhosphoSTAT2(Tyr690) l
46-624 Phospho-STAT3(Ser727) l l l
46-623 Phospho-STAT3(Tyr705) l l l
46-641 Phospho-STAT5A(Tyr694) l l l
46-633 Phospho-STAT6(Tyr641) l
46-661 Phospho-ZAP70(Tyr) l
Forthemostupdatedversionofthistable,visitwww.millipore.com/milliplex_csp.Thenclickthe“MAPmateBufferTable”link.
MILLIPLEX MAP Comparative Data
PerformancecharacteristicsofMILLIPLEXMAPCellSignalingMAPmateproductswere
benchmarkedagainstanalytesselectedfromtwocompetitors.Atotaloffive
phosphoproteinanalytes(Table1)wereselectedbasedonavailabilityfromMillipore
andcompetitorcompanies,aswellasmaximalsignalingpathwayoverlap.
60
0
pEGF
R
pERK
1/2
p-p70
S6K
pJNK/S
APK1
p-p38
80
100
40
20
Millipore Competitor A Competitor B
60
0pERK1/2 p-p70 S6K pJNK/SAPK1 p-p38
80
100
40
20
150
0pEGFR
200
250
100
50
Millipore Competitor A Competitor B
Comparison of Assaysusing Normalized Lysates
Comparison of Assays:Fold Induction
To determine fold induction the stimulated average MFI was divided by the unstimulated average MFI.
Protocol and Assay Design:oVerystraightforwardanduser-friendly
oNoadditionalsampletreatmentforsomeMAPmates
oOneprotocoldesignforallMAPmateproductsandkits
Kit Format: oAllkitsdesignedwiththesameformatformultiplexing
withtheleastoptimization
Buffers Required:oOnlytwobuffersrequired–DilutionandWashbuffers
Incubation Time and Temperature:
oOvernightincubationat4°C
oShorterincubationtimeatroomtemperaturepossible
Standards and Controls:oLyophilizednegativecontrol(unstimulatedlysate)
includedinMILLIPLEXMAPBufferandDetectionkit
oLyophilizedpositivecontrol(stimulatedlysate)included
withphospho-MAPmates
Advantages of MILLIPLEX MAP Cell Signaling Portfolio: Simpler, Easier Multiplexing
60
0
pEGF
R
pERK
1/2
p-p70
S6K
pJNK/S
APK1
p-p38
80
100
40
20
Millipore Competitor A Competitor B
60
0pERK1/2 p-p70 S6K pJNK/SAPK1 p-p38
80
100
40
20
150
0pEGFR
200
250
100
50
Millipore Competitor A Competitor B
Comparison of Assaysusing Normalized Lysates
Comparison of Assays:Fold Induction
To determine fold induction the stimulated average MFI was divided by the unstimulated average MFI.
Figure A and B.Manufacturer’sinstructionsforbothcelllysis,samplepreparationandforprotocolassayprocedureswerefollowedforeachofthecellsignalingkits(CatalogueNos.46-603,46-602,46-613,46-610,and46-629).Theresultsbelowshowingthefoldinductionvalues,aswellasfoldinductionwithnormalizedlysates,demonstratethesuperiorproductperformanceoftheMILLIPLEXMAPCellSignalingproductline.
Target Lysate Tested
pEGFR A431/EGF
pJNK/SAPK1 HeLa/HS-Ars
pERK1/2 A431/EGF
p-p38 HeLa/HS-Ars
p-p70S6K A431/EGF
Table 1.
A.
B.
xMAP TECHNOLOGY PROCESS FLOW
With xMAP technology, you will benefit from
the ability to simultaneously quantitate up to
100 analytes in a single sample, using as little
as 25 µL or less.
• Speed/High-Throughput: Simultaneously
measuretheconcentrationofalargenumberof
analytes,allowingyoutodomorefaster,gaining
earlyandcomprehensivedatasocriticaltoyour
work.
• Accuracy: xMAPtechnologygeneratesreal-time
analysisandaccuratequantificationofbiological
interactions.
• Reproducibility: Thehigh-volumeproduction
ofxMAPbeadsallowsassaystandardizationthat
solid-phasedflatarrayscannotprovide.
Luminex xMAP Technology
1 Luminexusesproprietarytechniquestointernallycolor-codemicrosphereswithtwofluorescentdyes.Throughpreciseconcentrationsofthesedyes,100distinctlycoloredbeadsetscanbecreated,eachofwhichiscoatedwithaspecificcaptureantibody.
3 Themicrospheresareallowedtopassrapidlythroughalaser,whichexcitestheinternaldyesmarkingthemicrosphereset.AsecondlaserexcitesPE,thefluorescentdyeonthereportermolecule.
2 Afterananalytefromatestsampleiscapturedbythebead,abiotinylateddetectionantibodyisintroduced.ThereactionmixtureisthenincubatedwithStreptavidinPEconjugate,thereportermolecule,tocompletethereactiononthesurfaceofeachmicrosphere.
Bead #12, IL-6Bead #29, IFN-γ
Bead #56, TNF-α
COMPARISON OF THE xMAP & WESTERN BLOT TECHNOLOGIESSide-by-sidecomparisonof
Multiplexingvs.WesternBlot
Traditional Method Multiplexing
“Western on a Bead”
Total Proteins
Phosphorylated Proteins
Western
1 Protein/probe
PRODUCTS • Higher Quality–Industry-leadingQC,QAandSixSigmaPrinciples.
• More Efficient–Analyticallyvalidatedkitsforconsistencyandreproducibility.
• All-in-One Kit–Asinglekitwithasinglecataloguenumber.
• More Flexible–Selectonlytheanalytesneededorchooseapre-mixedkit.
• Continuity of Knowledge Is the Key–Millipore“owns”everystepoftheprocess,
fromR&Dtomanufacturing.
• More Choices–ThebroadestrangeofmultiplexanalytesfortheLuminexxMAPplatform.
INSTRUMENTS • Speed/High-Throughput–Simultaneouslymeasuretheconcentrationofalargenumber
ofdifferentanalytes,allowingyoutodomorefaster,gainingearlyandcomprehensive
datasocriticaltoyourwork.
• Accuracy–xMAPtechnologygeneratesreal-timeanalysisandaccuratequantificationof
thebiologicalinteractions.
• Reproducibility–High-volumeproductionofxMAPmicrospheresallowsassay
standardizationthatsolid-phasedflatarrayscannotprovide.
• Low Sample Volume–Withminimalhands-ontime,youcanscreenmorethan
40analytesinasinglesampleusingaslittleas25µLorless.
SOFTWARE • TheLuminex xPONENT® softwarehelpsyoumanageandtrackallofyourmultiplex
assaysbycombiningeaseofuseandflexibilitytoperformbothproteinandnucleicacid
testingononesystem.It’salsocompatiblewiththeupcomingmagneticbead(MagPlex™
beads)andMILLIPLEXmagassays,andwillbemovingforwardwithcompatiblefuture
instrumentplatforms.
• NEW! MilliporenowoffersMILLIPLEX Analyst software,developedexclusivelyfor
MilliporebyVigeneTech®.Thisdataanalysispackagehelpsyoudoyourbestworkwith
increasedspeedandefficiency.MILLIPLEXAnalystsoftwareprovidesawholenewlevel
ofdataanalysisbyallowingyoutoimportlargedatasetsandbenefitfromtheabilityto
calculateyourresultsinseconds.Theuser-friendlyplatesetupfeatureletsyouwork
withspeedandefficiencywhiletheuniquealgorithmsoneventhemostchallengingdata
setsgiveyoutheperformanceyouneedformultiplexinganalysis.
SERVICES • Millipore’sBioMarkerServiceshasbeenperformingimmunoassaysforover25years.
• AssayservicesareavailableforRIAs,ELISAs,IRMAsandmultiplexassays,usingboth
Milliporekitsaswellasanycommerciallyavailableassays.
• Qualitycontrol,reproducibleresults,andconfidentialityareofparamountimportanceto
ourBioMarkerServicesteam.
• Millipore’sRegulatory Compliant Laboratoryisuniquelypositionedtoprovide
bioanalyticalservicestosupportyourpreclinicalandclinicalstudies.
MILLIPLEX MAP: Everything You Need—All In One Place
MILLIPLEX MAP
MILLIPLEX Analyzer
MILLIPLEX Analyst
BioPharma Services
Foracompleteandup-to-datelistofMILLIPLEXMAPanalytes,visitwww.millipore.com/analytes.
Description Catalogue No.
PhosphoAkt/PKB(Ser473),AssayBuffer1
46-601
PhosphoERK1/2(Thr185/Tyr187) 46-602
PhosphoEGFRec(Tyr) 46-603
ActiveCaspase3 46-604
TotalAkt/PKB,AssayBuffer1 46-605
TotalEGFReceptor 46-606
PhosphoHsp27(Ser78) 46-607
TotalHsp27 46-608
TotalERK1/2 46-609
Phosphop38/SAPK(Thr180/Tyr182)
46-610
PhosphoRsk1(Ser380) 46-611
Totalp38 46-612
PhosphoJNK/SAPK1(Thr183/Tyr185)
46-613
PhosphoPDGFReceptora(Tyr) 46-614
PhosphoPDGFReceptorb(Tyr) 46-615
PhosphoLck(Tyr) 46-616
TotalLck 46-617
TotalJNK 46-618
Phosphoc-Kit(Tyr) 46-619
Totalc-Kit 46-620
Totalp21 46-621
Phosphoc-Jun(Ser73) 46-622
PhosphoSTAT3(Tyr705) 46-623
PhosphoSTAT3(Ser727) 46-624
TotalSTAT3 46-625
PhosphoIRS1(Tyr) 46-627
TotalIRS1 46-628
Phosphop70S6Kinase(Thr412) 46-629
Totalp70S6Kinase 46-630
PhosphoCREB(Ser133) 46-631
TotalCREB 46-632
PhosphoSTAT6(Tyr641) 46-633
TotalSTAT2 46-634
PhosphoSTAT2(Tyr690) 46-635
PhosphoSTAT5A/B(Tyr694/Tyr699)
46-641
PhosphoIkBa(Ser32) 46-643
TotalIkBa 46-644
PhosphoAkt/PKB(Thr308),AssayBuffer1
46-645
Description Quantity Catalogue No.
Multi-Pathway8-Plex 96-wellplate 48-680
HumanTCellReceptor7-Plex 96-wellplate 48-690
HumanApoptosis3-Plex 96-wellplate 48-670
HumanMAPKinase9-Plex 96-wellplate 48-660
HumanSrcFamilyKinase-8-Plex 96-wellplate 48-650
HumanAkt/mTOR 96-wellplate 48-611
CellSignalingBufferandDetectionKit 96-wellplate 48-602
Description Catalogue No.
TotalIGF1R 46-646
PhosphoIGF1R(Tyr) 46-647
TotalErbB2/Her2 46-648
PhosphoErbB2/Her2(Tyr) 46-649
Totalc-Met/HGFR 46-650
Phosphoc-Met/HGFR(Tyr) 46-651
TotalSTAT1 46-654
PhosphoSTAT1(Tyr701) 46-655
CleavedPARP(Asp214) 46-656
TotalATF2 46-657
PhosphoATF2(Thr69/71) 46-658
TotalBcl-2 46-659
PhosphoBcl-2(Ser70) 46-660
PhosphoZAP-70(Tyr) 46-661
Totalp53 46-662
Phosphop53(Ser15) 46-663
Phosphop53(Ser46) 46-664
TotalHIF-1a 46-665
TotalHumanGAPDH 46-667
TotalMEK1 46-669
PhosphoMEK1(Ser222) 46-670
TotalMKK6 46-671
PhosphoMKK6(Ser207/Thr211) 46-672
TotalMSK1 46-673
PhosphoMSK1(Ser212) 46-674
TotalAkt/PKB,AssayBuffer2 46-675
PhosphoAkt/PKB(Thr308),AssayBuffer2
46-676
PhosphoAkt/PKB(Ser473),AssayBuffer2
46-677
TotalPTEN 46-678
PhosphoPTEN(Ser380) 46-679
TotalPRAS40 46-680
PhosphoPRAS40(Thr246) 46-681
TotalmTOR 46-685
PhosphomTOR(Ser2448) 46-686
TotalIR 46-687
PhosphoIR(Tyr1162/1163) 46-688
PhosphoHistoneH2A.X(Ser139) 46-692
PhosphoBad(Ser112 46-694
TotalBad 46-695
ORDERING INFORMATION
MILLIPLEX MAP MAPmate Products—96-well plate
MILLIPLEX MAP Cell Lysates—96-well plateDescription Catalogue No.
HeLa:unstimulated 47-205
Jurkat:unstimulated 47-206
Jurkat:anisomycin 47-207
Jurkat:H2O2 47-208
A431:EGF 47-210
HeLa:HS/Ars 47-211
HeLa:TNFa 47-212
NIH3T3:PDGF 47-213
Kasumi-3:SCF 47-214
Description Catalogue No.
HEK293:LiCl 47-215
MCF-7:IGF 47-216
Daudi:IL-4 47-217
A549:camptothecin 47-218
3T3:anisomycin 47-219
Jurkat:paclitaxel 47-220
3T3:unstimulated 47-221
HeLa:HGF 47-222
Other Related Millipore ProductsoMILLIPLEXmapandMILLIPLEXmagCytokine/ChemokineMultiplex
Panels:human,highsensitivityhuman,non-humanprimate,
mouse,rat,canine
oMILLIPLEXmapEpiQuantPanels:human
oMILLIPLEXAnalyst
oBioMarkerServices
oKinaseProfiler™ServicesandProducts
oGPCRProfiler®ServicesandProducts
oCellSignalingAntibodies
Millipore,AdvancingLifeScienceTogether,MILLIPLEX,andGPCRProfilerareregisteredtrademarksofMilliporeCorporation.EpiQuant,KinaseProfiler,PathwayProfiler,MAPmate,andtheMmarkaretrademarksofMilliporeCorporation.Luminex,xPONENT,andxMAPareregisteredtrademarksofLuminexCorporation.Luminex200andMagPlexisatrademarkofLuminexCorporation.VigeneTechisaregisteredtrademarkofVigeneTech,Inc.Lit.No.PB2723EN00,Rev.BPrintedinU.S.A.andFrance07/10LS-SBU-09-02583©2010MilliporeCorporation,Billerica,MA01821U.S.A.Allrightsreserved.
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