CatalogueNumber | FCCS100182 |
TradeName | |
Description | FlowCellect?HistoneH2A.XPhosphorylationAssayKit |
Overview | Millipore’sFlowCellect?HistoneH2A.XPhosphorylationAssaykitincludesonedirectlyconjugatedantibody,aphospho-specificAnti-phospho-HistoneH2A.X(Ser139)-AlexaFluor?488conjugatedantibodytomeasurelevelsofHistoneH2A.X.ThiskitisdesignedtodetecttheextentofHistoneH2A.XpathwayactivationbymeasuringH2A.Xphosphorylationinanygivencellpopulation.ThekitprovidesasensitiveandvaluabletooltostudythefactorsthatinduceDNAdamageand/oraffectDNArepair,andallowonetoexplorethelinkagebetweenDNAdamage,cellcyclecheckpoints,andinitiationofapoptosis.
IonizingrADIation(IR)andmanychemotherapeuticagentslikeetoposidekillcancercellsbyinductionofDNADSBs.Severalreportsshowthatthelevelofgamma-H2A.XasdetectedbyflowcytometrycorrelateswiththenumberofDNAstrandbreaks,tothelevelofcelldeathandradiosensitivity.H2A.XphosphorylatesinresponsetoaDNAdamagingreagent(e.g.Etoposide)orUVlight,anditsactivationclearlyindicatesthatDSBshaveoccurred.UnderstandingwhenDSBstakeplacecanhelpresearchersunderstandthemechanismsinvolvedinDNArepairandtheDNAdamageresponse.
TheantibodyprovidedinthekithasbeencarefullytitratedtoensuretheABIlitytomeasurephospho-HistoneH2A.Xforaccuratedeterminationofproteinactivation.Sufficientreagentsareprovidedtoperform100tests.Detailedassayinstructionsareincludedtoassistinanalysisandtoensurethecorrectcellconcentrationisobtainedduringacquisitionofsampledata. |
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BackgroundInformation | H2A.XisamemberofthehistoneH2Afamily.HistoneH2A.XresidesdownstreamoftheDNAdamagekinasesignalingcascade.PhosphorylationofHistoneH2A.Xatserine139isanimportantindicatorofDNAdamage.AsthelevelofDNAdamageincreases,thelevelofphosphoHistoneH2A.X(alsoknownasγH2AX)increases,accumulatingatthesitesofDNAdamage.ThisaccumulationofphosphoHistoneH2A.XisoftenusedtoindicatethelevelofDNAdamagepresentwithinthecell.H2AXisalsoresponsIBLeforrecruitingresponseproteinstothesiteofDNAdamageandmayplayaroleinDNArepair.
Inall,acomprehensiveunderstandingofHistoneH2A.XactivityandunderstandingtheconsequenceofitsactivationcanprovidetheresearcherwithusefulinformationwhichwillbeimportantinunderstandingtheintrinsicnatureoftheDNAdamageresponse.Millipore’sFlowCellectHistoneH2A.XPhosphorylationAssayKitisdesignedtoallowtheresearchertomonitorandaccuratelymeasurephospho-specificHistoneH2A.Xactivationinapopulationofcells.
AllFlowCellectkitsareoptimizedonguava?benchtopflowcytometers.FlowCellectkitscanbeusedonanyflowcytometerfollowingthesameprotocolprovidingresearchersareliableandfullyvalidatedsolutiontostudytheHistoneH2A.Xsignalingpathwayrightinthecomfortoftheirownlab.Thedirectlyconjugatedantibodyprovidedinthekithasbeencarefullytitratedandoptimizedtoensuremaximalperformancealleviatingtheneedforanyadditionaloptimization.Thiskitcontainsoptimizedfixation,permeabilization,wash,andassaybufferstoprovideresearcherswithacompletesolutionforHistoneH2A.Xsignalinganalysis. |