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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
CatalogueNumber	17-488
BrandFamily	Upstate
TradeName	STAR Upstate
Description	phospho-p38α(Thr180/Tyr182)STARELISAKit
BackgroundInformation	I.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.
MaterialsRequiredbutNotDelivered	1.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.
Detectionmethod	Colorimetric

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.

Detectionmethod

Colorimetric

StorageandShippingInformation
StorageConditions	Maintaintheunopenedkitat2-8°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.

StorageandShippingInformation

StorageConditions

Maintaintheunopenedkitat2-8°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
Application	Thisphospho-p38α(Thr180/Tyr182)STARELISAKitisusedtomeasure&quantifyp38/SAPKlevels.
KeyApplications	ELISA

BIOLOGicalInformation
SpeciesReactivity	Human
AnalytesAvailable	p38/SAPK
EntrezGeneNumber	NM_001315.1 NM_139012.1 NM_139013.1 NM_139014.1
EntrezGeneSummary	TheproteinencodedbythisgeneisamemberoftheMAPkinasefamily.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	Q16539
UniProtSummary	FUNCTION: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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Millipore/17-488 | phospho-p38α (Thr180/Tyr182) STAR ELISA Kit/17-488/96 assays

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