Millipore/FCCS025100 | FlowCellect™ PI3K/MAPK Dual Pathway Activation and Cancer Marker Detection kit/FCCS025100/25 Test,Millipore,,Millipore,密理博,merckmillipore,Millipore公司代理生物计划密理博产品目录价格,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：Millipore/FCCS025100 | FlowCellect™ PI3K/MAPK Dual Pathway Activation and Cancer Marker Detection kit/FCCS025100/25 Test

产品编号：FCCS025100

市场价：¥10400.00

场地：美国(厂家直采)

产品分类：试剂盒>ELISA试剂盒>夹心法ELISA>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：$520.00

品牌： Millipore

公司：Millipore

公司分类：

商品介绍

Description
CatalogueNumber	FCCS025100
TradeName	FlowCellect
Description	FlowCellect™PI3K/MAPKDualPathwayActivationandCancerMarkerDetectionkit
Overview	Recentevidencesuggeststhatcross-talkbetweenthePI3KandMAPKsignalingpathwaysexist.WeusepAktandpERKantibodiestoexaminethePI3K/MAPKinteractions.Additionally,tofurtherinterrogatetheinterplaybetweenthesetwopathways,cellproliferativemarkerKi-67isusedtovalidatethefinalBIOLOGicaleffect. Millipore’sFlowCellect™PI3K/MAPKDualActivationandCancerDetectionKitisdesignedtoexaminethiscross-talkinamulti-parametricfashionbyprovidingthreefullyvalidatedandoptimizedantibodybiomarkerstomeasurespecificcellsignalingeventsinflowapplications.ThethreeantibodiesprovidedinthekitareAnti-phospho-AktAlexaFluor488conjugate,Anti-phospho-ERKR-Phycoerythrinconjugate,andAnti-Ki-67PerCPconjugate.Byutilizingallthreeantibodybiomarkerssimultaneouslyinflowapplications,wenowhavetheABIlitytothoroughlyevaluatethe“cross-talk”betweenPI3KandMAPKpathwaysandtofurtherdeterminetheconsequenceoftheirinterplayincellproliferationanddifferentiationbymeasuringtheireffectonKi-67expression. PhosphorylatedAktandphosphorylatedERKareincludedinthekittoprovidetheenduserwiththemeanstocross-examineboththePI3KandMAPKsignalingpathwayssimultaneously.IthasbeensuggestedthatthephosphorylationofAktcanresultintheinhibition,ordephosphorylation,ofphospho-RafonSer259[Jun,T.etal.(1999)].ByinactivatingRaf,thiswillessentiallyblocktheMAPKsignalingpathwayresultinginaninactivatedphospho-ERK.Insomesituations,asurfacereceptor(suchasIGF-1)willactivateboththePI3KandMAPKpathwaysleADIngtothephosphorylationofbothAktandERK.However,sincethisinteractionor“cross-talk”existbetweenthetwopathways,itiscriticaltoinvestigatetheirinteractionsinbothaspatialandtemporalmanner. Inarecentstudy,wehaveexaminedtheeffectsofIGF-1activationbytheadditionofInsulinonboththePI3KandMAPKsignalingpathwaysonHEK293cells.Wehaveperformedthisexperimentimplementingcriticaltimepointsforsignalingevaluation:activationat3minuteandat5minutetimeintervals.Theresultingresponsesindicatethatcross-talkisobserved,notedbyasharpdecreaseinERKexpression.ThistransientresponseisattributedtothephosphorylationofAkt,whichinturnwillshutoffphosphorylatedERKasnotedabove. Inordertovalidatethebiologicaleffectofphospho-proteinactivationbyagivenstimulus,cellcyclemarkerKi-67isusedsinceitisatruemeasurementofthe“proliferativefraction”.Ki-67ispresentinallphasesofthecellcycleexceptforG0.However,Ki-67canonlybedetectedinflowcytometrywhencellsaregoingthroughMphaseasKi-67expressionpatternsarepunctateinallotherphasesproducingweakersignals.Butbyusingacellcyclearrestreagent,CellCycleStop™,cellproliferationmeasuredbyKi-67expressioncanbeaccuratelydeterminedascellsarearrestedatMphaseandareclearlyvisIBLeinflowcytometryanalysis.InordertoaccuratelymeasurethecellproliferativeactivitybyKi-67expression,however,cellsmustbetreatedforatleast12hourswithacombinationofCellCycleStop™andagivencellstimulustofullyachieveenoughcirculatingcellstobecapturedinM-phase.Additionally,priortocelltreatmentsallculturesmustbeserumstarvedfor24hourstoessentiallyresetthecellcycleandbringmostcirculatingcellsbacktoG0[Littleton,RJ.etal.(1991)]. Usingmulti-parametricflowanalysis,weareabletocross-examinethesesignalingeventsandtheirbiologicalconsequencesimultaneously,providingabiologicalcorrelationbetweenpathwayactivationandcancerproliferation.
AlternateNames	FlowCellect
BackgroundInformation	Examinationofcellsignalingpathwaysandmonitoringtheiractivationstatushavebeenextremelyimportantforresearcherstounderstandthedetailedmechanismsofcellularfunctionsandthecauseofvariousdiseases.Manysignaltransductionpathwayshavebeenimplicatedtoleadtomultipleoutcomessuchasapoptosis,celldifferentiation,cellgrowthandcellproliferation,allofwhichhavebeenextensivelystudiedforthetreatmentofvariouscancersandautoimmunediseases. Thestudyofcellsignalingpathwaysarenowmadeeasierwiththeuseofactivationstatus-specificandphospho-specificantibodies.Measurementofproteinphosphorylationwithphospho-specificantibodieshasgiveninsightintokinasesignalingcascades[Krutzik,P.O.etal.(2003)].Multi-parameterphosphoflowcytometryisapowerfultoolforstudyingmultiplepathwaysinamixedcellpopulationatthesametime. Muchexcitementinthefieldofsignaltransductionhascenteredonthediscoveryofincreasingcross-talkamongsignalingpathways[Jun,T.etal.(1999)].RecentevidencehassuggestedthatcommunicationbetweenthePI3KandMAPKpathwaysexistdownstreamfromthecellsurface[Jun,T.etal.(1999),Moelling,K.etal.(2002),Zimmermann,S.etal.(1999)].Theabilityforsignalingpathwaystocross-talkaddsanextradimensionandcomplexitywhenevaluatingpathwaysofinterest.Sincesignaltransductionpathwaysareanelaboratehighwayofevents,theabilitytomonitorthesekeyintracellular“checkpoints”simultaneouslyprovidesresearchersaverypowerfultoolforanalyzingcomplicatedcelleventssuchascancercellproliferationbymeasuringtheactivityofmultiplecellsignalingpathways. Millipore’sFlowCellect™PI3K/MAPKDualPathwayActivationandCancerMarkerDetectionkitisdesignedtoallowtheresearchertocrossexamineboththePI3KandMAPKsignalingpathways,aswellascellproliferationsimultaneously.Thiskitprovidesthreedirectlyconjugatedantibodieswhichareoptimizedformulti-colorflowcytometryapplicationsforthedetectionofAktphosphorylation,ERK1/2phosphorylationandKi-67cancermarkerexpression.Ki-67hasbeenindicatedtobeareliabletumorproliferativemarkerincancercells[Ishikuro,A.etal.(1997)].ThefractionofKi-67positivecells,oftendefinedasthe“proliferativefraction”,hasprognosticvalueinmanytumors[Darzynkiewicz,Z.etal.(2001)]. Althoughtheuseofphospho-specificantibodystainingasabiomarkermaygivesomemeasureoftargetactivation,itmaynotnecessarilycorrelatewiththedesiredbiologicaleffect(e.g.growthinhibitionorapoptosis).Researcherswouldlikelybenefitfromtheinclusionofbothphospho-specificsignaltransductionmarkers(suchaspERKandpAkt)andaproliferationmarker(suchasKi-67)toallowatruemeasureoftreatmentevaluationatthelevelofthetumor[Smalley,KSMetal.(2007)]. AllFlowCellect™kitsareoptimizedonthebench-topGuava®flowcytometrysystems,whichsavesvaluabletimeandsamplevolume.Allkitscontainoptimizedfixation,permeabilization,washandflowbufferstoprovideresearcherswithacompletesolutionforsimultaneousdetectionofmultiplepathwayactivations.WiththeGuavaplatformandFlowCellect™kits,onecanfinallyhaveaneasy,reliableandfullyvalidatedsolutiontostudythecomplexcellsignalingpathwaysrightinthecomfortofyourownlab.

ProductInformation
Components	1.20XAnti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)R-PhycoerythrinconjugateMonoclonalAntibody:(PartNo.CS203329)One150µLvial 2.20XAnti-phospho-Akt1/PKBα(Ser473)AlexaFluor488conjugateMonoclonalAntibody:(PartNo.CS203326)One150µLvial 3.20XAnti-KI-67PerCPconjugateMonoclonalAntibody:(PartNo.CS203324)One150µLvial 4.FixationBuffer:(PartNo.CS202122)One13mLbottle 5.10XWashBuffer:(PartNo.CS202123)One13mLbottle 6.5XAssayBuffer:(PartNo.CS202124)One55mLbottle 7.1XPermeabilizationBuffer:(PartNo.CS202125)One13mLbottle 8.CellCycleStop™:(PartNo.CS203335)Onevialsuppliedwith1mg
Detectionmethod	¹²⁵IFluorescent

ProductInformation

Components

1.20XAnti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)R-PhycoerythrinconjugateMonoclonalAntibody:(PartNo.CS203329)One150µLvial
2.20XAnti-phospho-Akt1/PKBα(Ser473)AlexaFluor488conjugateMonoclonalAntibody:(PartNo.CS203326)One150µLvial
3.20XAnti-KI-67PerCPconjugateMonoclonalAntibody:(PartNo.CS203324)One150µLvial
4.FixationBuffer:(PartNo.CS202122)One13mLbottle
5.10XWashBuffer:(PartNo.CS202123)One13mLbottle
6.5XAssayBuffer:(PartNo.CS202124)One55mLbottle
7.1XPermeabilizationBuffer:(PartNo.CS202125)One13mLbottle
8.CellCycleStop™:(PartNo.CS203335)Onevialsuppliedwith1mg

Detectionmethod

¹²⁵IFluorescent

StorageandShippingInformation
StorageConditions	4-8°Cforantibodiesandbuffers

Applications
Application	ThisFlowCellectPI3K/MAPKDualActivation&CancerDetectionKitisdesignedtoexaminethiscross-talkinamulti-parametricfashionbyproviding3validatedbiomarkerstomeasurespecificcellsignalingeventsinflowcytometryapplications.
KeyApplications	FlowCytometry

BiologicalInformation
Host	Mouse
SpeciesReactivity	Human
AntibodyType	MonoclonalAntibody

PhysicochemicalInformation

Dimensions

MaterialsInformation

MaterialsInformation

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