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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
CatalogueNumber	17-472
BrandFamily	Upstate
TradeName	STAR Upstate
Description	Phospho-GSK-3β(Ser9)STARELISAKit
AlternateNames	glycogensynthasekinase3beta
BackgroundInformation	I.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.
MaterialsRequiredbutNotDelivered	1.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.
Detectionmethod	Colorimetric

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.

Detectionmethod

Colorimetric

StorageandShippingInformation
StorageConditions	Maintaintheunopenedkitat2-8°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.

StorageandShippingInformation

StorageConditions

Maintaintheunopenedkitat2-8°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
Application	ThisPhospho-GSK-3β(Ser9)STARELISAKitisusedtomeasure&quantifyGSK-3βlevels.
KeyApplications	ELISA

BIOLOGicalInformation
SpeciesReactivity	Human Mouse Rat
AnalytesAvailable	GSK-3β
EntrezGeneNumber	NM_002093.2
EntrezGeneSummary	Glycogensynthasekinase-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	P49841
UniProtSummary	FUNCTION: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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Millipore/17-472 | Phospho-GSK-3β (Ser9) STAR ELISA Kit/17-472/96 assays

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