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    當前位置 ： Millipore >>> Millipore/17-488 | phospho-p38α (Thr180/Tyr182) STAR ELISA Kit/17-488/96 assays
    Millipore/17-488 | phospho-p38α (Thr180/Tyr182) STAR ELISA Kit/17-488/96 assays
    • Millipore/17-488 | phospho-p38α (Thr180/Tyr182) STAR ELISA Kit/17-488/96 assays

    Millipore/17-488 | phospho-p38α (Thr180/Tyr182) STAR ELISA Kit/17-488/96 assays

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    貨號: 17-488
    品牌: Millipore
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      • Description
        CatalogueNumber17-488
        BrandFamilyUpstate
        TradeName
        • STAR
        • Upstate
        Descriptionphospho-p38α(Thr180/Tyr182)STARELISAKit
        BackgroundInformationI.TESTPRINCIPLE
        TheUPSTATE?colorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.Thep38MAPKplateiscoatedwithaspecificmousemonoclonalp38MAPKcaptureantibodyonthemicrowellsofthe96-wellclearplate.Samplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingp38MAPKantigentobecapturedintheplatewells.Theplateisthenwashedtoremoveanynon-boundunspecificmaterial.Thewellsarethenincubatedwithaspecificrabbitanti-phospho-p38(Thr180/Tyr182)antibodytodetectthecapturedp38(Thr180/Tyr182)ontheplatewell.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.Thisallowsforasensitiveenzymaticdetectionofthesample.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader.
        Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.Thekitalsoincludesastandardthatisrunasbothapositivecontrolandtodevelopastandardlineofdetection.

        II.p38BACKGROUND
        Thep38MAPKfamilyofSer/Thrkinaseproteinsismadeupof4isoforms,p38alpha(MAPK14andSAPK2a),p38beta(MAPK11andSAPK2b),p38delta(MAPK13,SAPK4),andp38gamma(MAPK12,SAPK3).Theotherp38isoformsshare74%,57%,and60%sequenceidentitywiththealphaisoform,respectively.AllfourisoformspossestheTGYmotifandareactivatedbythedualphosphorylationofthethreonineandtyrosineresiduesinthismotif,whichcorrespondtoThr180/Tyr182inhumanp38alpha.p38,especiallythealphaandbetaisoforms,areabletophosphorylateawidearrayoftargetsincludingnumerouskinasesincludingMAPKAP-K2,MAPKAP-K3,PRAK,MSK,andMNKaswellastranscriptionfactorsincludingSTAT1,p53,NFATp,ATF-2,Elk-1(althoughweakly),andMEF2a/c.p38MAPKfamilymembershavetrADItionallybeenassociatedwithstress,immuneresponse,andregulationofcellproliferationandapoptosis.Theyalsohavearoleinregulatingtheproductionofpro-inflammatorycytokinesandhavebeenassociatedwithdiseasessuchasasthmaandseveralautoimmunediseases,includingrheumatoidarthritis,diabetes,andinflammatoryboweldisease.JustasERKisphosphorylatedandactivatedbyMEK,p38alphaMAPKinaseisactivatedpredominantlybyitsupstreamregulatorykinasesMKK6andMKK3aswellasMKK4whichalsoactivatesJNK,butisnotasubstrateforMKK7likeJNKis.
        MaterialsRequiredbutNotDelivered1.Multi-channelorrepeatingPipettes
        2.Plateshaker(optional)
        3.Pipettors&tipscapableofaccuratelymeasuring1-1000μL
        4.GraduatedSEROlogicalpipettes
        5.96-wellmicrotiterPlateReaderwith450nmfilter
        6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata
        7.Microfugetubesforstandardandsampledilutions
        8.Mechanicalvortex
        9.1litercontainer
        10.Distilledordeionizedwater
        ProductInformation
        Components
        • CapturePlatepre-coatedwithanti-p38alphaMAPKantibody:(PartNo.17-487A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
        • Anti-phospho-p38alpha(Thr180/Tyr182)detectionantibody:(PartNo.17-487B)Onebottle(11mL)ofanti-phospho-p38alpha(Thr180/Tyr182)detectionantibodycontainingsodiumazide,readytouse.
        • ELISADiluent:(PartNo.17-487C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
        • 25XELISAWashBuffer:(PartNo.17-487D)Onebottle(50mL)of25XELISAWashBuffer.
        • Anti-RabbitIgGHRPconjugate:(PartNo.17-487E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
        • HRPDiluent:(PartNo.17-487F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
        • TMBSolution:(PartNo.17-487G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
        • StopSolution:(PartNo.17-487H)Onebottle(25mL)ofstopsolution,readytouse.
        • phospho-p38alpha(Thr180/Tyr182)Standard:(PartNo.17-487I)Twovialsofphospho-p38alpha(Thr180/Tyr182)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-p38α(Thr180/Tyr182)STARELISAKitisusedtomeasure&quantifyp38/SAPKlevels.
        KeyApplications
        • ELISA
        BIOLOGicalInformation
        SpeciesReactivity
        • Human
        AnalytesAvailable
        • p38/SAPK
        EntrezGeneNumber
        EntrezGeneSummaryTheproteinencodedbythisgeneisamemberoftheMAPkinasefamily.MAPkinasesactasanintegrationpointformultiplebiochemicalsignals,andareinvolvedinawidevarietyofcellularprocessessuchasproliferation,differentiation,transcriptionregulationanddevelopment.Thiskinaseisactivatedbyvariousenvironmentalstressesandproinflammatorycytokines.TheactivationrequiresitsphosphorylationbyMAPkinasekinases(MKKs),oritsautophosphorylationtriggeredbytheinteractionofMAP3K7IP1/TAB1proteinwiththiskinase.ThesubstratesofthiskinaseincludetranscriptionregulatorATF2,MEF2C,andMAX,cellcycleregulatorCDC25B,andtumorsuppressorp53,whichsuggesttherolesofthiskinaseinstressrelatedtranscriptionandcellcycleregulation,aswellasingenotoxicstressresponse.Fouralternativelysplicedtranscriptvariantsofthisgeneencodingdistinctisoformshavebeenreported.
        GeneSymbol
        • MAPK14
        • PRKM14
        • RK
        • SAPK2A
        • CSBP1
        • p38
        • CSPB1
        • CSBP
        • PRKM15
        • Mxi2
        • p38ALPHA
        • MXI2
        • EXIP
        • CSBP2
        Modifications
        • Phosphorylation
        UniProtNumber
        UniProtSummaryFUNCTION:SwissProt:Q16539#Respondstoactivationbyenvironmentalstress,pro-inflammatorycytokinesandlipopolysaccharide(LPS)byphosphorylatinganumberoftranscriptionfactors,suchasELK1andATF2andseveraldownstreamkinases,suchasMAPKAPK2andMAPKAPK5.Playsacriticalroleintheproductionofsomecytokines,forexampleIL-6.MayplayaroleinstabilizationofEPOmRNAduringhypoxicstress.IsoformMxi2activationisstimulatedbymitogensandoxidativestressandonlypoorlyphosphorylatesELK1andATF2.IsoformExipmayplayaroleintheearlyonsetofapoptosis.
        COFACTOR:Magnesium.
        SIZE:360aminoacids;41293Da
        SUBUNIT:Bindstoakinaseinteractionmotifwithintheproteintyrosinephosphatase,PTPRR.ThisinteractionretainsMAPK14inthecytoplasmandpreventsnuclearaccumulation.InteractswithSPAG9(Bysimilarity).InteractswithNP60.
        SUBCELLULARLOCATION:Cytoplasm(Bysimilarity).Nucleus(Bysimilarity).
        TISSUESPECIFICITY:Brain,heart,placenta,pancreasandskeletalmuscle.Expressedtoalesserextentinlung,liverandkidney.DOMAIN:SwissProt:Q16539TheTXYmotifcontainsthethreonineandtyrosineresidueswhosephosphorylationactivatestheMAPkinases.
        PTM:DuallyphosphorylatedonThr-180andTyr-182,whichactivatestheenzyme.&PhosphorylateduponDNAdamage,probablybyATMorATR.
        SIMILARITY:Belongstotheproteinkinasesuperfamily.CMGCSer/Thrproteinkinasefamily.MAPkinasesubfamily.&Contains1proteinkinasedomain.
        PhysicochemicalInformation
        Sensitivity
        • Sensitivity:1Unit/mL.
          RangeofDetection:1.6to100Units/mL
          SpeciesReactivity:Human
        Dimensions
        MaterialsInformation
        MaterialsInformation
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