Pierce/ERK1/ERK2 (Phospho) [pT185/pY187] Human ELISA Kit/96 tests/KHO0091

Description

TheERK1/2[pTpY185/187]ELISAresearchuseonlykitisdesignedtodetectandquantifythelevelofERK2proteinphosphorylatedatthreonineresidue185andtyrosine187andERK1atthreonine202andtyrosine204.ThisassayisintendedforthedetectionofERK1/2[pTpY185/187]fromhumancelllysatesusing96-wellplatesandamicroplatereader.

•Easytorun
•Moresensitivethanwesterns
•Convenientprecoated,removable8-wellstrips

Easy-to-RunSandwichELISA
Thetotalassayincubationtimeisonly4hours.AmonoclonalcaptureantibodyspecificforERK1/2hasbeencoatedontothewellsofthe96-wellplateprovided.Duringthefirstincubation,standardsofknownERK1/2[pTpY185/187]contentandunknownsamplesarePipettedintothewellsandtheERK1/2antigenbindstotheimmobilized(capture)antibody.Afterwashing,arabbitantibodyspecificforERK1/2[pTpY185/187]isaddedtothewells.Duringthesecondincubation,thisantibodyservesasadetectionantibodybybindingtotheimmobilizedERK1/2proteincapturedduringthefirstincubation.Afterwashing,ahorserADIshperoxidase–labeledanti–rabbitIgGisadded.Thisbindstothedetectionantibodytocompletethefour–membersandwich.Afterathirdincubationandwashingtoremovealltheunboundenzyme,asubstratesolution(TMB)isadded,whichisacteduponbytheboundenzymetoproducecolor.TheintensityofthiscoloredproductisdirectlyproportionaltotheconcentrationofERK1/2[pTpY185/187]presentintheoriginalspecimenandtheopticaldensitycanbereadonastandardmicroplatereader.

DetectLow-ExpressingProteinsandUseLessSample
ELISAkitsdesignedtomeasureintracellularsignalingtargetsare2–10timesmoresensitivethanwesternblotting.Theimprovedsensitivityenablesyoutodetectlow-expressingproteinsthatotherwisemaynotbedistinguishablefrombackground.Inaddition,theamountofsampleneededtoruntheassayislessthanwhatisneededforwesternblots.

Ready-to-UseERK1/2[pTpY185/187]ELISAKitContainsTheseReagents,Buffers,andSupplies:
•ERK1/2AntibodyCoated96-WellPlate
•ERK1/2[pTpY185/187]Standard
•StandardDiluentBuffer
•ERK1/2[pTpY185/187]DetectionAntibody
•Anti-RabbitIgG-HRP(100X)
•HRPDiluent
•WashBufferConcentrate(25X)
•StABIlizedChromogen,TMB
•StopSolution
•PlateCovers
•Detailedprotocolwithvalidationtests

ConvenientKitFormat
Thekitcomeswithacaptureantibodyprecoatedtotheplate,eliminatinganovernightcoatingprocess.Theplatesaremachinecoatedtoprovidelowvariabilitywell-to-well.FurThermore,theplatecomesasremovable8-wellstripsallowingyoutorunasfewsamplesasyouwant.Inaddition,thestandardsprovidedinthekitcanbeusedtogetquantitativeresultsoffthestandardcurveand/orcanbeusedasapositivecontrol.

RigorousValidation
EachELISAkitisvalidatedforsensitivity,specificity,precision,andlot-to-lotconsistency.Seeprotocolinsertformoreinformationonvalidation.

PartoftheELISAKitFamily
WehaveawidemenuofELISAresearchuseonlykits.Withexcellentsensitivity,provenspecificity,andconsistentperformance,theseoptimizedELISAkitsofferyouachoicewhenitcomestogettingphysiologicallyrelevantandquantitativeresults.

Forresearchuseonly.Notintendedforhumanoranimaltherapeuticordiagnosticuse.

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