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

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

Millipore 提供三种不同的 Immobilon PVDF 膜，是针对不同的蛋白质印迹应用的理想选择。 现在我们还 提供采用按规格裁切的印迹方片膜和两层过滤纸组成的印迹三明治。   与硝酸纤维素膜相比，Immobilon 膜具有多种优点。 它们不易破裂或卷 曲，便于裁剪且在切割过程中不会碎裂。 它们还有低背景、广泛溶剂相容性以及优异的着色能力。 此外，它们可以重复检测。 Immobilon-P 膜（沃比森-供应） ·         0.45μm 孔径 ·         建议用于大多数免疫印迹，特别在蛋 白质大于 20kDa 时 ·         Millipore 的快速免疫检测实验方法不需要膜封闭并缩短洗涤时间，将检测时间至少减少了两个小时，而不致降低灵敏 度或特异性 用于高通量实验室的印迹三明治 ·         当您需要处理多个印迹时，印迹三明治是一种便利省时的选择。 我们的印迹三明 治包含多片 Immobilon-P 膜，其中插入了按规格裁切的多张层析级印迹纸。 转印滤纸 ·         按照大多数免疫印迹尺寸进行裁切的层析级转印滤纸。 Immobilon-PSQ 膜（沃比森-供应） ·         0.2μm 孔径和大的内表面面积 ·         较其它膜具有更高的 蛋白质吸附率和测序得率 ·         建议用于小于 20kDa 的印迹蛋白质 ·         防止小分子量的蛋白质的渗漏 Immobilon-FL 膜（沃比森-供应） ·        针对荧光应用进行优化的转印膜 ·         极低的背景提高了所 有荧光检测实验的灵敏度 ·         与所有常用的荧光探针（所有激发和发射波长）兼容 ·         是多态性检测和化学荧光应用的理想选择