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℡ 4000-520-616
℡ 4000-520-616
Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays
产品编号:17-472
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场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
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美  元  价:待定
品      牌: Millipore
公      司:Millipore
公司分类:
Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays
商品介绍
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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