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    當前位置 ： Millipore >>> Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays
    Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays
    • Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays

    Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays

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    貨號: 17-472
    品牌: Millipore
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      • Description
        CatalogueNumber17-472
        BrandFamilyUpstate
        TradeName
        • STAR
        • Upstate
        DescriptionPhospho-GSK-3β(Ser9)STARELISAKit
        AlternateNames
        • glycogensynthasekinase3beta
        BackgroundInformationI.TESTPRINCIPLETheUPSTATE?colorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheGSK-3βplateiscoatedwithaspecificmousemonoclonalGSK-3&betacaptureantibodyonthemicrowellsofthe96-wellclearplate.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingGSK-3βantigentobecapturedintheplatewells.Theplateisthenwashedtoremoveanynon-boundunspecificmaterial.Thewellsarethenincubatedwithaspecificrabbitanti-GSK-3βantibodytodetectthecapturedphosphor-GSK-3β(Ser9)ontheplatewell.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.Thisallowsforasensitiveenzymaticdetectionofthesample.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader.Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.Thekitalsoincludesastandardthatisrunasbothapositivecontrolandtodevelopastandardlineofdetection.

        II.GSK-3βBACKGROUNDGlycogenSynthaseKinase-3differsfrommostserine/threoninekinasesinthatitisactiveintheabsenceoftheactionofsignalingpathways.Whenitbecomesphosphorylated,theproteinbecomesinactive.Therearetwoisoformsthatexist,GSK-3αandGSK-3β.ThefunctionofGSK-3istophosphorylateGlycogenSynthaseandtherebyinactivateit.InsulinactionstimulatesthePI3KinasepathwaythatresultsintheactivationofAkt.AktphosphorylatesGSK-3,therebyinactivatingit.GlycogenSynthase,GSK-3"sdownstreamsubstrate,isthenrapidlydephosphorylatedandactivated.OtherGSK-3substratesincludeJun(oninhibitorysites),andeIF2B.DetectionofGSK-3"sphosphorylationstatusoftheAktsite(Ser21)onGSK-3issuitableforsurrogateassaysoftheactivationstateofthepathway.
        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
        • GSK-3βCapturePlate:(PartNo.17-472A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
        • Anti-phospho-GSK-3β(Ser9)detectionantibody:(PartNo.17-472B)Onebottle(6mL)ofanti-phospho-GSK-3β(Ser9)detectionantibodycontainingsodiumazide,readytouse.
        • ELISADiluent:(PartNo.17-472C)Onebottle(25mL)ofELISADiluentsodiumazide,readytouse
        • 25XELISAWashBuffer:(PartNo.17-472D)Onebottle(50mL)of25XELISAWashBuffer.
        • Anti-RabbitIgGHRPconjugate:(PartNo.17-472E)Onevial(125μL)of100Xanti-rabbitHRPconjugate.
        • HRPDiluent:(PartNo.17-472F)Onebottle(25mL)ofHRPdiluent.
        • TMBSolution:(PartNo.17-472G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
        • StopSolution:(PartNo.17-472H)Onebottle(25mL)ofstopsolution,readytouse.
        • PhosphorylatedGSK-3β(Ser9)Standard:(PartNo.17-472I)TwovialsofphosphorylatedGSK-3β(Ser9)standard,lyophilized.
        • PlateCovers:Twoplatecovers.
        DetectionmethodColorimetric
        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
        ApplicationThisPhospho-GSK-3β(Ser9)STARELISAKitisusedtomeasure&quantifyGSK-3βlevels.
        KeyApplications
        • ELISA
        BIOLOGicalInformation
        SpeciesReactivity
        • Human
        • Mouse
        • Rat
        AnalytesAvailable
        • GSK-3β
        EntrezGeneNumber
        EntrezGeneSummaryGlycogensynthasekinase-3(GSK3)isaproline-directedserine-threoninekinasethatwasinitiallyidentifiedasaphosphorylatingandinactivatingglycogensynthase.Twoisoforms,alpha(GSK3A;MIM606784)andbeta,showahighdegreeofaminoacidhomology(StambolicandWoodgett,1994[PubMed7980435]).GSK3Bisinvolvedinenergymetabolism,neuronalcelldevelopment,andbodypatternformation(Plyteetal.,1992[PubMed1333807]).[suppliedbyOMIM]
        GeneSymbol
        • GSK3B
        Modifications
        • Phosphorylation
        UniProtNumber
        UniProtSummaryFUNCTION:SwissProt:P49841#ParticipatesintheWntsignalingpathway.Implicatedinthehormonalcontrolofseveralregulatoryproteinsincludingglycogensynthase,MYBandthetranscriptionfactorJUN.PhosphorylatesJUNatsitesproximaltoitsDNA-bindingdomain,therebyreducingitsaffinityforDNA.
        SIZE:420aminoacids;46744Da
        SUBUNIT:Monomer(Bysimilarity).InteractswithNINandCABYR.
        TISSUESPECIFICITY:Expressedintestis,thymus,prostateandovaryandweaklyexpressedinlung,brainandkidney.
        PTM:PhosphorylatedbyAKT1andILK1.
        SIMILARITY:SwissProt:P49841##Belongstotheproteinkinasesuperfamily.CMGCSer/Thrproteinkinasefamily.GSK-3subfamily.&Contains1proteinkinasedomain.
        PhysicochemicalInformation
        Sensitivity
        • Sensitivity:1.6Units/mL
          RangeofDetection:1.6to100Units/mL
        Dimensions
        MaterialsInformation
        MaterialsInformation
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