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

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貨號: 17-465

品牌: Millipore

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Description
CatalogueNumber	17-465
BrandFamily	Upstate
TradeName	STAR Upstate
Description	JNK1/2STARELISAKit
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
BackgroundInformation	testPrinciple 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.
MaterialsRequiredbutNotDelivered	1.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.
Detectionmethod	Chromogenic

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.

Detectionmethod

Chromogenic

StorageandShippingInformation
StorageConditions	1yearat4°Cfromdateofshipment

Applications
Application	TheJNK1/2colorimetricSTARELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtoquantifyspecificlevelsofsignalingtargetsindenaturedcellextracts.
KeyApplications	ELISA

BIOLOGicalInformation
SpeciesReactivity	Human Mouse
AnalytesAvailable	JNK1/2
EntrezGeneNumber	NM_002750.2 NM_139046.1 NM_139047.1 NM_139049.1
EntrezGeneSummary	TheproteinencodedbythisgeneisamemberoftheMAPkinasefamily.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	P45983 P45984
UniProtSummary	FUNCTION: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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