Description
CatalogueNumber	FCCS025106
TradeName	FlowCellect
Description	FlowCellect™MAPKActivationDualDetectionKit
Overview	Millipore’sFlowCellect™MAPKActivationDualDetectionkitincludestwodirectlyconjugatedantibodies,aphospho-specificAnti-phospho-ERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PEandanAnti-ERK1/2-AlexaFluor®647conjugatedantibodytomeasuretotallevelsofERK.ThistwocolorflowcytometrykitisdesignedtomeasuretheextentofMAPKphosphorylationrelativetothetotalMAPKexpressioninanygivencellpopulation.Bydoingsuch,thelevelsofboththetotalandphosphorylatedproteincanbemeasuredsimultaneouslyinthesamecell,resultinginanormalizedandaccuratemeasurementofMAPKactivationafterstimulation.Moreover,simultaneousmeasurementofbothtotalandphospho-ERK1/2confirmstargetspecificityofthephosphorylationevent.Together,atotalandphosphoantibodyduoperformedinmultiplexprovidesanenhancedandmorereliabledetectionofthephospho:totalratiowithinamixedpopulation. AllFlowCellectkitsaresuppliedwithoptimizedfixation,permeABIlization,wash,andassaybufferstoprovideresearcherstheabilitytodetecttheactivationofERKinflowapplications.Sufficientreagentsareprovidedfor25two-colorsamples.Thekitincludesalloptimizedfluorescentlylabeledantibodiesandbuffersnecessaryforcellpreparationandanalysis.
AlternateNames	FlowCellect
BackgroundInformation	Erk(Extracellularsignal-RelatedKinase)isafamilyoftwo,highlyhomologousproteinsdenotedasErk1(p44,MAPK3)andErk2(p42,MAPK1)thatbothfunctioninthesamepathway.ThetwoproteinsareoftenreferredtocollectivelyasErk1/2orp44/p42MAPkinase.TheErkpathwayisconsideredtheclassical,canonicalMAPK(Mitogen-ActivatedProteinKinase)signalingpathway.Itisanevolutionarilyconservedpathwaythatcontrolsandisacriticalregulatorofthegrowthandsurvivalthroughthepromotionofcellproliferationandthepreventionofapoptosis.Erkisinvolvedinthecontrolofmanyfundamentalcellularprocessesincludingcellproliferation,survival,differentiation,apoptosis,motilityandmetabolism.Erkisactivatedbygrowthfactorstimulationofreceptortyrosinekinases(RTKs),GPCR,and/orintegrinstimulation.ThisactivatestheRas-Raf-MEK-Erkpathwaythatresultsinthephosphorylation/activationofErk1/2(p44/p42)ontheTxYmotif(Thr202/Tyr204andThr185/Tyr187forErk1&Erk2,respectively). Theregulationofalargenumberofcellularprocesses(suchascellproliferation/differentiation,cellsurvivalandapoptosis)isdependentontheactivationofMAPKsignalingpathway,whichcanbemeasuredbythephosphorylationofERK1/2.

ProductInformation
Components	1.20XAnti-phospho-ERK1/2(Thr202/Tyr204,Thr185/Tyr187)PEConjugatedAntibody:(PartNo.CS203329)Onevialcontaining150µL. 2.20XAnti-ERK1/2-AlexaFluor®647ConjugatedAntibody:(PartNo.CS205075)Onevialcontaining150µL. 3.FixationBuffer:(PartNo.CS202122)Onebottlecontaining13mL. 4.10XWashBuffer:(PartNo.CS202123)Onebottlecontaining13mL. 5.5XAssayBuffer:(PartNo.CS202124)One55mLbottle 6.1XPermeabilizationBuffer:(PartNo.CS203284)Onebottlecontaining13mL.
Detectionmethod	Fluorescent

StorageandShippingInformation
StorageConditions	4-8°Cforantibodiesandbuffers

Applications
Application	ThisFlowCellectMAPKActivationDualDetectionKitforFlowCytometryincludes2conjugatedantibodies,phospho-specificAnti-phospho-ERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE&Anti-ERK1/2-AlexaFluor647tomeasuretotallevelsofERK.
KeyApplications	FlowCytometry

BIOLOGicalInformation
Host	Mouse
SpeciesReactivity	Human
AntibodyType	MonoclonalAntibody

PhysicochemicalInformation

Dimensions

MaterialsInformation

MaterialsInformation

Millipore 提供三种不同的 Immobilon PVDF 膜，是针对不同的蛋白质印迹应用的理想选择。 现在我们还 提供采用按规格裁切的印迹方片膜和两层过滤纸组成的印迹三明治。   与硝酸纤维素膜相比，Immobilon 膜具有多种优点。 它们不易破裂或卷 曲，便于裁剪且在切割过程中不会碎裂。 它们还有低背景、广泛溶剂相容性以及优异的着色能力。 此外，它们可以重复检测。 Immobilon-P 膜（沃比森-供应） ·         0.45μm 孔径 ·         建议用于大多数免疫印迹，特别在蛋 白质大于 20kDa 时 ·         Millipore 的快速免疫检测实验方法不需要膜封闭并缩短洗涤时间，将检测时间至少减少了两个小时，而不致降低灵敏 度或特异性 用于高通量实验室的印迹三明治 ·         当您需要处理多个印迹时，印迹三明治是一种便利省时的选择。 我们的印迹三明 治包含多片 Immobilon-P 膜，其中插入了按规格裁切的多张层析级印迹纸。 转印滤纸 ·         按照大多数免疫印迹尺寸进行裁切的层析级转印滤纸。 Immobilon-PSQ 膜（沃比森-供应） ·         0.2μm 孔径和大的内表面面积 ·         较其它膜具有更高的 蛋白质吸附率和测序得率 ·         建议用于小于 20kDa 的印迹蛋白质 ·         防止小分子量的蛋白质的渗漏 Immobilon-FL 膜（沃比森-供应） ·        针对荧光应用进行优化的转印膜 ·         极低的背景提高了所 有荧光检测实验的灵敏度 ·         与所有常用的荧光探针（所有激发和发射波长）兼容 ·         是多态性检测和化学荧光应用的理想选择