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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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货号: 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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