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Millipore/17-465 | JNK 1/2 STAR ELISA Kit/17-465/96 assays

Millipore/17-465 | JNK 1/2 STAR ELISA Kit/17-465/96 assays

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貨號(hào): 17-465
品牌: Millipore
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    • Description
      CatalogueNumber17-465
      BrandFamilyUpstate
      TradeName
      • STAR
      • Upstate
      DescriptionJNK1/2STARELISAKit
      AlternateNames
      • Mitogen-activatedproteinkinase8
      • MAPkinase8
      • MAPK8
      • JNK-46
      • Stress-activatedproteinkinase1c
      • SAPK1c
      • Stress-activatedproteinkinaseJNK1
      • (c-JunN-terminalkinase1
      • Mitogen-activatedproteinkinase9
      • MAPkinase9
      • MAPK9
      • JNK-55
      • Stress-activatedproteinkinase1a
      • SAPK1a
      • Stress-activatedproteinkinaseJNK2
      • c-JunN-terminalkinase2
      BackgroundInformationtestPrinciple
      TheUPSTATEcolorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheJNK1/2plateiscoatedwithaspecificmousemonoclonalJNK1/2captureantibodyonthemicrowellsofthe96-wellclearplate.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingJNK1/2antigentobecapturedintheplatewells.Theplateisthenwashedtoremoveanynon-boundunspecificmaterial.Thewellsarethenincubatedwithaspecificrabbitanti-JNK1/2antibodytodetectthecapturedJNKontheplatewell.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-channelorrepeatingPipettes2.Plateshaker(optional)3.Pipettors&tipscapableofaccuratelymeasuring1-1000μL4.GraduatedSEROlogicalpipettes5.96-wellmicrotiterPlateReaderwith450nmfilter6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata7.Microfugetubesforstandardandsampledilutions8.Mechanicalvortex9.1litercontainer10.Distilledordeionizedwater
      ProductInformation
      Components
      • 1.CapturePlatepre-coatedwithanti-JNK1/2antibody:(PartNo.17-465A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
      • 2.Anti-JNK1/2detectionantibody:(PartNo.17-465B)Onebottle(11mL)ofanti-JNK1/2detectionantibodycontainingsodiumazide,readytouse.
      • 3.ELISADiluent:(PartNo.17-465C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
      • 4.25XELISAWashBuffer:(PartNo.17-465D)Onebottle(50mL)of25XELISAWashBuffer.
      • 5.Anti-RabbitIgGHRPconjugate:(PartNo.17-465E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
      • 6.HRPDiluent:(PartNo.17-465F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
      • 7.TMBSolution:(PartNo.17-465G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
      • 8.StopSolution:(PartNo.17-465H)Onebottle(25mL)ofstopsolution,readytouse.
      • 9.JNK1/2Standard:(PartNo.17-465I)TwovialsofJNK1/2standard,lyophilized.
      • 10.PlateCovers:Twoplatecovers.
      DetectionmethodChromogenic
      StorageandShippingInformation
      StorageConditions1yearat4°Cfromdateofshipment
      Applications
      ApplicationTheJNK1/2colorimetricSTARELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,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
      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:0.16ng/mL.RangeofDetection:0.31to20ng/mL
      Dimensions
      MaterialsInformation
      MaterialsInformation
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