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主营:分子类,蛋白类,抗体类,生化类试剂
℡ 4000-520-616
℡ 4000-520-616
Millipore/17-479 | STAR FAK ELISA Kit/17-479/96 assays
产品编号:17-479
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场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
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品      牌: Millipore
公      司:Millipore
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Millipore/17-479 | STAR FAK ELISA Kit/17-479/96 assays
商品介绍
Description
CatalogueNumber17-479
BrandFamilyUpstate
TradeName
  • STAR
  • Upstate
DescriptionSTARFAKELISAKit
AlternateNames
  • Focaladhesionkinase1
  • FADK1
  • Focaladhesionkinase-relatednonkinase
  • FRNK
  • Proteinphosphatase1regulatorysubunit71
  • PPP1R71
  • Protein-tyrosinekinase2
  • p125FAK
  • pp125FAK
BackgroundInformationI.TESTPRINCIPLE
TheUPSTATE®colorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheFAKplateiscoatedwithaspecificmousemonoclonalFAKcaptureantibody.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingFAKantigentobecaptured.Theplateisthenwashedtoremoveanyun-boundnon-specificmaterial.Aspecificrabbitanti-FAKantibodyisaddedtodetectthecapturedFAKontheplate.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader.Thisallowsforasensitiveenzymaticdetectionofthesample.
Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.ThekitalsoincludesastandardthatisrunasbothapositivecontrolandtogenerateastandardcurveforFAKmeasurement.

II.FAKBACKGROUND
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.
MaterialsRequiredbutNotDelivered1.Multi-channelorrepeatingPipettes
2.Plateshaker(optional)
3.Pipettorsandtipscapableofaccuratelymeasuring1-1000µL
4.GraduatedSEROlogicalpipettes
5.96-wellmicrotiterPlateReaderwith450nmfilter
6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata
7.Microfugetubesforstandardandsampledilutions
8.Mechanicalvortex
9.1litercontainer
10.Distilledordeionizedwater
ProductInformation
Components
  • CapturePlatepre-coatedwithanti-FAKantibody:(PartNo.17-479A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
  • Anti-FAKdetectionantibody:(PartNo.17-479B)Onebottle(11mL)ofanti-FAKdetectionantibodycontainingsodiumazide,readytouse.
  • ELISADiluent:(PartNo.17-479C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
  • 25XELISAWashBuffer:(PartNo.17-479D)Onebottle(50mL)of25XELISAWashBuffer.
  • Anti-RabbitIgGHRPconjugate:(PartNo.17-479E)Onevial(125µL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
  • HRPDiluent:(PartNo.17-479F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
  • TMBSolution:(PartNo.17-479G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
  • StopSolution:(PartNo.17-479H)Onebottle(25mL)ofstopsolution,readytouse.
  • FAKStandard:(PartNo.17-479I)TwovialsofFAKstandard,lyophilized.
  • PlateCovers:Twoplatecovers.
DetectionmethodChromogenic
StorageandShippingInformation
StorageConditionsMaintaintheunopenedkitat2-8°Cuntilexpirationdate.

Precautions

•Theinstructionsprovidedhavebeendesignedtooptimizethekit"sperformance.Deviationfromtheinstructionsmayresultinsuboptimalperformanceofthekitandthefailuretoproduceaccuratedata.
•CausticMaterial:StopSolution.Caution:Eye,hand,face,andclothingprotectionshouldbewornwhenhandlingthismaterial.
•SafetyWarningsandPrecautions:Thiskitisdesignedforresearchuseonlyandnotrecommendedforinternaluseinhumansoranimals.Allchemicalsshouldbeconsideredpotentiallyhazardousandprinciplesofgoodlaboratorypracticeshouldbefollowed.
•TheDetectionAntibodyandELISADiluentcontainsodiumazide.Sodiumazidemayreactwithcopperandleadplumbingtoformhighlyexplosivemetalazides.Upondisposal,flushwithlargeamountsofwatertopreventazidebuild-up.Avoidcontactwithskin.
•TheAnti-RabbitIgGHRPConjugateandHRPDiluentcontainthimerosal.Thimerosalishighlytoxicbyinhalation,contactwithskinorifswallowed.Thimerosalisapossiblemutagenandshouldbehandledaccordingly.
Applications
ApplicationThisSTARFAKELISAKitisusedtomeasure&quantifyFAKlevels.
KeyApplications
  • ELISA
BiologicalInformation
SpeciesReactivity
  • Human
  • Mouse
  • Rat
AnalytesAvailable
  • FAK
EntrezGeneNumber
EntrezGeneSummaryThisgeneencodesacytoplasmicproteintyrosinekinasewhichisfoundconcentratedinthefocaladhesionsthatformbetweencellsgrowinginthepresenceofextracellularmatrixconstituents.TheencodedproteinisamemberoftheFAKsubfamilyofproteintyrosinekinasesbutlackssignificantsequencesimilaritytokinasesfromothersubfamilies.Activationofthisgenemaybeanimportantearlystepincellgrowthandintracellularsignaltransductionpathwaystriggeredinresponsetocertainneuralpeptidesortocellinteractionswiththeextracellularmatrix.Atleastfourtranscriptvariantsencodingfourdifferentisoformshavebeenfoundforthisgene,butthefull-lengthnaturesofonlytwoofthemhavebeendetermined.
GeneSymbol
  • PTK2
  • pp125FAK
  • FADK
  • FAK1
  • FAK
UniProtNumber
UniProtSummaryFUNCTION: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:1.6ng/mL
    RangeofDetection:1.6to100ng/mL
    SpeciesReactivity:Human,mouseandrat
Dimensions
MaterialsInformation
MaterialsInformation
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