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Millipore/17-480 | Phospho-FAK (Tyr397) STAR ELISA Kit/17-480/96 assays

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貨號: 17-480

品牌: Millipore

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Description
CatalogueNumber	17-480
BrandFamily	Upstate
TradeName	STAR Upstate
Description	Phospho-FAK(Tyr397)STARELISAKit
AlternateNames	Focaladhesionkinase1 FADK1 Focaladhesionkinase-relatednonkinase FRNK Proteinphosphatase1regulatorysubunit71 PPP1R71 Protein-tyrosinekinase2 p125FAK pp125FAK
BackgroundInformation	FAK(FocalAdhesionKinase)isanon-receptortyrosinekinasewhichlocalizestofocaladhesionsandplaysakeyroleinintegrin-mediatedsignalingpathwaysregulatingcellmigration.Itautophosphorylatesontyrosine397duringadhesionandspreADIng,andisthoughttobeacriticaltransducerofadhesion-dependentgrowthandsurvival.FAKanditphosphorylationstateshavebeenimplicatedincancermetastasisandtumorcellsurvivalandadhesion-independentgrowth.Additionally,recentevidenceindicatesthatelevationofFAKactivityinhumancarcinomacellsisassociatedwithincreasedinvasivepotential.AcentralroleintumorformationandprogressionsuggestthatFAKisanattractivetargetfortherapeuticintervention.Cellsurfaceintegrinswhenengagedwiththeirligandsleadtotherecruitmentofanumberofintracellularproteinstospecializedsitesofthecytoplasmicfaceintofocaladhesions.Thesefocaladhesionplaquesarecomposedofmanyproteinsthatincludekinases,phosphatases,scaffoldingproteins,andG-proteins.Theactivationofintegrinsleadstothetyrosinephosphorylationofseveralcytoplasmicproteinscriticaltothebiochemicalprocess,includingFAK,Src,paxillin,tensin,andp130Cas.Thisclusteringandactivationofproteinsatthemembraneleadstotheactivationofmanydivergingsignalingpathwaysthatresultincytoskeletalre-organization.ThisstartswitheithertherecruitmentofSrcandFAKtotheplasmamembranefollowingintegrinorPI3Kinaseactivation.Thenon-receptorprotein-tyrosinekinases(PTKs)FAKandSrcarekeymembersofthefocaladhesioncomplex.ThesePTKsco-Localizeandassociatewiththetransmembraneintegrinsandvariousdownstreamtargetsofcertaingrowthfactorreceptors.TheoutcomeofSrcbindingtoFAKisthephosphorylationofseveraltyrosineresiduesinFAKanditsrelatedproteins,includingtheFAKautophosphorylationsiteTyr397.ThisautophosphorylationoccursimmediatelyafterintegrinclusteringandallowsfortheassociationofFAKwithvarioussignalingproteinssuchasPI3kinase,Grb2-Sos,p130Cas,andpaxillin.ThephosphorylationofFAKTyr397iscrucialformanyoftheestablishedBIOLOGicalrolesofFAK,includingcellmigration,cellcycleprogression,andpreventionofanoikis,aformofdetachment-inducedapoptosisandiscorrelatedwiththeinvasivenessoftumorsandisthoughttobeapivotalplayerbymodulatingtheturnoveroffocaladhesionstoregulatecellmigration.Phosphoinositide3-Kinase(PI3kinaseorPI3K)alsoplaysaroleincellmigrationviaitsbindingtotheautophosphorylationsite(Tyr397)ofFAKthroughoneorbothofitsSH2domainsinthep85subunitofPI3K.ThisisthesamesiteinFAKthatisboundbySrc.Intheory,thebindingofPI3kinasetoFAKmaybeinvolvedinactivitieslikecellproliferation,apoptosis,andmigrationthatarerelatedtothephosphorylationofTyr397.ThisissubstantiatedbytheobservationthatAKTisdownstreamofPI3kinaseandisamediatorinpreventingapoptosis.ThereforeitispossIBLethatFAK/PI3kinaseassociationmayhelpregulateapoptosis.
MaterialsRequiredbutNotDelivered	1.Multi-channelorrepeatingPipettes2.Plateshaker(optional)3.Pipettorsandtipscapableofaccuratelymeasuring1-1000μL4.GraduatedSEROlogicalpipettes5.96-wellmicrotiterPlateReaderwith450nmfilter6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata7.Microfugetubesforstandardandsampledilutions8.Mechanicalvortex9.1litercontainer10.Distilledordeionizedwater

ProductInformation
Components	1.CapturePlatepre-coatedwithanti-FAKantibody:(PartNo.17-480A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch. 2.Anti-phospho-FAK(Tyr397)detectionantibody:(PartNo.17-480B)Onebottle(6mL)ofanti-phospho-FAK(Tyr397)detectionantibodycontainingsodiumazide,readytouse. 3.ELISADiluent:(PartNo.17-480C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse. 4.25XELISAWashBuffer:(PartNo.17-480D)Onebottle(50mL)of25XELISAWashBuffer. 5.Anti-RabbitIgGHRPconjugate:(PartNo.17-480E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal. 6.HRPDiluent:(PartNo.17-480F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal. 7.TMBSolution:(PartNo.17-480G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse. 8.StopSolution:(PartNo.17-480H)Onebottle(25mL)ofstopsolution,readytouse. 9.phospho-FAK(Tyr397)Standard:(PartNo.17-480I)Twovialsofphospho-FAK(Tyr397)standard,lyophilized. 10.PlateCovers:Twoplatecovers.
Detectionmethod	Chromogenic

ProductInformation

Components

1.CapturePlatepre-coatedwithanti-FAKantibody:(PartNo.17-480A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
2.Anti-phospho-FAK(Tyr397)detectionantibody:(PartNo.17-480B)Onebottle(6mL)ofanti-phospho-FAK(Tyr397)detectionantibodycontainingsodiumazide,readytouse.
3.ELISADiluent:(PartNo.17-480C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
4.25XELISAWashBuffer:(PartNo.17-480D)Onebottle(50mL)of25XELISAWashBuffer.
5.Anti-RabbitIgGHRPconjugate:(PartNo.17-480E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
6.HRPDiluent:(PartNo.17-480F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
7.TMBSolution:(PartNo.17-480G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
8.StopSolution:(PartNo.17-480H)Onebottle(25mL)ofstopsolution,readytouse.
9.phospho-FAK(Tyr397)Standard:(PartNo.17-480I)Twovialsofphospho-FAK(Tyr397)standard,lyophilized.
10.PlateCovers:Twoplatecovers.

Detectionmethod

Chromogenic

StorageandShippingInformation
StorageConditions	1yearat4°Cfromdateofshipment

Applications
Application	ThisPhospho-FAK(Tyr397)STARELISAKitisusedtomeasure&quantifyFAKlevels.
KeyApplications	EnzymeAssays

BiologicalInformation
SpeciesReactivity	Human Mouse Rat
AnalytesAvailable	FAK
EntrezGeneNumber	NM_005607.3 NM_153831.2
EntrezGeneSummary	Thisgeneencodesacytoplasmicproteintyrosinekinasewhichisfoundconcentratedinthefocaladhesionsthatformbetweencellsgrowinginthepresenceofextracellularmatrixconstituents.TheencodedproteinisamemberoftheFAKsubfamilyofproteintyrosinekinasesbutlackssignificantsequencesimilaritytokinasesfromothersubfamilies.Activationofthisgenemaybeanimportantearlystepincellgrowthandintracellularsignaltransductionpathwaystriggeredinresponsetocertainneuralpeptidesortocellinteractionswiththeextracellularmatrix.Atleastfourtranscriptvariantsencodingfourdifferentisoformshavebeenfoundforthisgene,butthefull-lengthnaturesofonlytwoofthemhavebeendetermined.
GeneSymbol	PTK2 pp125FAK FADK FAK1 FAK
Modifications	Phosphorylation
UniProtNumber	Q05397
UniProtSummary	FUNCTION:SwissProt:Q05397#Non-receptorprotein-tyrosinekinaseimplicatedinsignalingpathwaysinvolvedincellmotility,proliferationandapoptosis.Activatedbytyrosine-phosphorylationinresponsetoeitherintegrinclusteringinducedbycelladhesionorantibodycross-linking,orviaG-proteincoupledreceptor(GPCR)occupancybyligandssuchasbombesinorlysophosphatidicacid,orviaLDLreceptoroccupancy.Playsapotentialroleinoncogenictransformationsresultinginincreasedkinaseactivity. SIZE:1052aminoacids;119233Da SUBUNIT:InteractswithCASfamilymembersandwithGIT1,SORBS1andBCAR3.InteractswithSHB(Bysimilarity).InteractswithTGFB1I1. SUBCELLULARLOCATION:Celljunction,focaladhesion;Cytoplasmicside.Note=Constituentoffocaladhesions. TISSUESPECIFICITY:Expressedinallorganstested,inlymphoidcelllines,butmostabundantlyinbrain. DOMAIN:SwissProt:Q05397ThefirstPro-richdomaininteractswiththeSH3domainofCRK-associatedsubstrate(BCAR1)andCASL(Bysimilarity).&Thecarboxy-terminalregionisthesiteoffocaladhesiontargeting(FAT)sequencewhichmediatesthelocalizationofFAK1tofocaladhesions. PTM:Phosphorylatedon6tyrosineresiduesuponactivation. SIMILARITY:Belongstotheproteinkinasesuperfamily.Tyrproteinkinasefamily.FAKsubfamily.&Contains1FERMdomain.&Contains1proteinkinasedomain.

PhysicochemicalInformation
Sensitivity	Sensitivity:1.6Units/mL. RangeofDetection:1.6to100Units/mL

Dimensions

MaterialsInformation

MaterialsInformation

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