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微孔/FCCS025100 | Flowcell等™ PI3K/MAPK双通路激活和肿瘤标志物检测试剂盒/FCCS025100/25试验
微孔/FCCS025100 | Flowcell等™ PI3K/MAPK双通路激活和肿瘤标志物检测试剂盒/FCCS025100/25试验
商品编号：
FCCS025100
品牌：
密理博
市场价：
¥10400.00
美元价：
6240.00
产品分类：
夹心法ELISA
公司分类：
Sandwich_method_ELISA
联系Q Q：
3392242852
电话号码：
4000-520-616
电子邮箱：
info@ebiomall.com
商品介绍
| Description | |
|---|---|
| CatalogueNumber | FCCS025100 |
| TradeName |
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| 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. |
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| 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 | |
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| Components |
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| Detectionmethod | 125IFluorescent |
| StorageandShippingInformation | |
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| StorageConditions | 4-8°Cforantibodiesandbuffers |
| BiologicalInformation | |
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| Host | Mouse |
| SpeciesReactivity |
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| AntibodyType | MonoclonalAntibody |
| PhysicochemicalInformation |
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| Dimensions |
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| MaterialsInformation |
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| MaterialsInformation |
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品牌介绍
密理博(Millipore)公司成立于1954年，总部位于美国麻省，在全世界设有47个办事处，为100多个国家提供产品和技术服务。目前全球雇员超过5800人，在美国、法国和日本等国家拥有7家大型生产工厂，主要生产过滤膜及膜过滤产品。20世纪80年代，密理博公司进入中国市场。先后在香港、北京、上海、广州及成都设立了办事机构，并于2000年4月在上海浦东外高桥保税区建立了密理博（上海）贸易有限公司。为了更好地满足中国用户的需求，密理博中国主页于2006年11月向广大用户开放，介绍密理博中国有限公司的最新动态，力求为用户打造专业的产品与服务信息交流平台。
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