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    當前位置 ： Millipore >>> Millipore/17-466 | phospho-JNK 1/2 (Thr183/Tyr185) STAR ELISA Kit/17-466/96 assays
    Millipore/17-466 | phospho-JNK 1/2 (Thr183/Tyr185) STAR ELISA Kit/17-466/96 assays
    • Millipore/17-466 | phospho-JNK 1/2 (Thr183/Tyr185) STAR ELISA Kit/17-466/96 assays

    Millipore/17-466 | phospho-JNK 1/2 (Thr183/Tyr185) STAR ELISA Kit/17-466/96 assays

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    貨號: 17-466
    品牌: Millipore
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      • Description
        CatalogueNumber17-466
        BrandFamilyUpstate
        TradeName
        • STAR
        • Upstate
        Descriptionphospho-JNK1/2(Thr183/Tyr185)STARELISAKit
        BackgroundInformationTestPrinciple
        TheUPSTATEcolorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsindenaturedcellextracts.TheJNK1/2plateiscoatedwithaspecificmousemonoclonalJNK1/2captureantibodyonthemicrowellsofthe96-wellclearplate.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingJNK1/2antigentobecapturedintheplatewells.Theplateisthenwashedtoremoveanynon-boundunspecificmaterial.Thewellsarethenincubatedwithaspecificrabbitanti-phospho-JNK(Thr183/Tyr185)antibodytodetectthecapturedJNKontheplatewellthathasbeenphosphorylatedonThr183andTyr185.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.Thisallowsforasensitiveenzymaticdetectionofthesample.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader.Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.Thekitalsoincludesastandardthatisrunasbothapositivecontrolandtodevelopastandardlineofdetection.

        JNKBackground
        JNK(c-JunN-terminalKinase,MAPK8)isamemberoftheMAPkinasefamily.JNKwasoriginallyfoundtobestressactivated,andassucharealsoreferredtoasthestress-relatedMAPK.FouralternativelysplicedtranscriptvariantsencodingdistinctisoformsofJNKhavebeenreported.JNKisregulatedbystressinducingsignalssuchasosmoticstressandUVrADIationaswellasproinflammatorycytokinessuchasIL-1andTNFα.JustasErk(p44/p42,MAPK)isactivatedbyitsupstreamkinaseMEK,JNKisphosphorylatedandactivatedbyMKK4andMKK7onthequintessentialMAPKactivationsequenceofThr-X-Tyr,whereXisaProfortheJNKkinasesonThr183/Tyr185forJNK1andThr221/Tyr223forJNK2.Oncephosphorylatedandactivated,JNKgoesontophosphorylatedownstreamtargetsinboththecytoplasmandthenucleus.JNKservesasatranscriptionalregulatorandisknowntophosphorylateanumberofdownstreamtargets,includingthetranscriptionfactorsc-Jun,ATF-2,andELK-1.C-Jun,amemberoftheAP-1familyoftranscriptionfactors,regulatescytokinegeneexpression.Alongtheselines,JNKinhibitorsmaybeeffectiveinthecontrolofrheumatoidarthritis(RH),anautoimmunediseaseinvolvingincreasedproductionofinflammatorycytokines.InhibitionofJNKsignalingalsoenhanceschemotherapy-inducedinhibitionoftumorcellgrowth,suggestingtheJNKsmayprovideamoleculartargetforthetreatmentofcancer.BothJNKandp38MAPKareoftensimultaneouslyactivatedinresponsetoenvironmentalchanges.Thesetwostresskinasesignalingpathwayshaveevolvedtorelayincreasinglycomplexrangesofenvironmentalstimuli.TheJNKpathwayiscriticallyinvolvedindiabetesandlevelsareabnormallyelevatedinobesity.Thecell-permeableJNKinhibitorypeptidemayhavepromiseasatherapeuticagentfordiabetes.
        MaterialsRequiredbutNotDelivered1.Multi-channelorrepeatingPipettes
        2.Plateshaker(optional)
        3.Pipettors&tipscapableofaccuratelymeasuring
        1-1000μL
        4.GraduatedSEROlogicalpipettes
        5.96-wellmicrotiterPlateReaderwith450nmfilter
        6.Graphingsoftwareforplottingdataorgraphpaperformanual
        plottingofdata
        7.Microfugetubesforstandardandsampledilutions
        8.Mechanicalvortex
        9.1litercontainer
        10.Distilledordeionizedwater
        ProductInformation
        Components
        • 1.CapturePlatepre-coatedwithanti-JNK1/2antibody:(PartNo.17-466A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
        • 2.Anti-phospho-JNK(Thr183/Tyr185)detectionantibody:(PartNo.17-466B)Onebottle(11mL)ofanti-phospho-JNK(Thr183/Tyr185)detectionantibodycontainingsodiumazide,readytouse.
        • 3.ELISADiluent:(PartNo.17-466C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
        • 4.25XELISAWashBuffer:(PartNo.17-466D)Onebottle(50mL)of25XELISAWashBuffer.
        • 5.Anti-RabbitIgGHRPconjugate:(PartNo.17-466E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
        • 6.HRPDiluent:(PartNo.17-466F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
        • 7.TMBSolution:(PartNo.17-466G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
        • 8.StopSolution:(PartNo.17-466H)Onebottle(25mL)ofstopsolution,readytouse.
        • 9.Phospho-JNK(Thr183/Tyr185)Standard:(PartNo.17-466I)TwovialsofphosphorylatedJNK(Thr183/Tyr185)standard,lyophilized.
        • 10.PlateCovers:Twoplatecovers.
        DetectionmethodChromogenic
        StorageandShippingInformation
        StorageConditions1yearat4°Cfromdateofshipment
        Applications
        ApplicationThephospho-JNK1/2(Thr183/Tyr185)colorimetricSTARELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtoquantifyspecificlevelsofsignalingtargetsindenaturedcellextracts.
        KeyApplications
        • ELISA
        BIOLOGicalInformation
        SpeciesReactivity
        • Human
        • Mouse
        AnalytesAvailable
        • JNK1/2
        EntrezGeneNumber
        EntrezGeneSummaryTheproteinencodedbythisgeneisamemberoftheMAPkinasefamily.MAPkinasesactasanintegrationpointformultiplebiochemicalsignals,andareinvolvedinawidevarietyofcellularprocessessuchasproliferation,differentiation,transcriptionregulationanddevelopment.Thiskinasetargetsspecifictranscriptionfactors,andthusmediatesimmediate-earlygeneexpressioninresponsetovariouscellstimuli.ItismostcloselyrelatedtoMAPK8,bothofwhichareinvolvedinUVradiationinducedapoptosis,thoughttoberelatedtothecytochromec-mediatedcelldeathpathway.ThisgeneandMAPK8arealsoknownasc-JunN-terminalkinases.Thiskinaseblockstheubiquitinationoftumorsuppressorp53,andthusitincreasesthestabilityofp53innonstressedcells.Studiesofthisgene"smousecounterpartsuggestakeyroleinT-celldifferentiation.Fouralternativelysplicedtranscriptvariantsencodingdistinctisoformshavebeenreported.
        GeneSymbol
        • MAPK8
        • JNK1
        • JNK21B1/2
        • PRKM8
        • JNK-46
        • JNK
        • SAPK1
        • JNK1A2
        Modifications
        • Phosphorylation
        UniProtNumber
        UniProtSummaryFUNCTION:SwissProt:P45984#JNK2isoformsdisplaydifferentbindingpatterns:alpha-1andalpha-2preferentiallybindtoc-Jun,whereasbeta-1andbeta-2bindtoATF2.However,thereisnocorrelationbetweenbindingandphosphorylation,whichisachievedataboutthesameefficiencybyallisoforms.JUNBisnotasubstrateforJNK2alpha-2,andJUNDbindsonlyweaklytoit.
        COFACTOR:Magnesium.
        SIZE:424aminoacids;48139Da
        SUBUNIT:Bindstoatleastfourscaffoldingproteins,MAPK8IP1/JIP-1,MAPK8IP2/JIP-2,MAPK8IP3/JIP-3/JSAP1andSPAG9/MAPK8IP4/JIP-4.TheseproteinsalsobindothercomponentsoftheJNKsignalingpathway.
        DOMAIN:SwissProt:P45984TheTXYmotifcontainsthethreonineandtyrosineresidueswhosephosphorylationactivatestheMAPkinases.
        PTM:DuallyphosphorylatedonThr-183andTyr-185,whichactivatestheenzyme.Autophosphorylatedinvitro.
        SIMILARITY:Belongstotheproteinkinasesuperfamily.CMGCSer/Thrproteinkinasefamily.MAPkinasesubfamily.&Contains1proteinkinasedomain.
        PhysicochemicalInformation
        Sensitivity
        • Sensitivity:0.8unit/mL.
          RangeofDetection:1.6to100units/mL
          SpeciesReactivity:Humanandmouse
        Dimensions
        MaterialsInformation
        MaterialsInformation
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