Viafluor™ SECellProliferationKitsuseamine-reactivedyestocovalentlylabelthecellcytoplasm,andcanbeusedtomonitorcelldivisionbyflowcytometry.
Themembranepermeablecompoundisnon-fluorescentuntilitentersviablecells,whereitishydrolyzedbycytoplasmicesteraseenzymestoreleasesthefluorescentamine-reactivedye.Thedyesthencovalentlyreactwithaminegroupsonintracellularproteins,formingfluorescentconjugatesthatareretainedinthecell,whileexcessunreacteddyeiswashedaway. Immediatelyafterstaining,asingle,brightfluorescentpopulationwillbedetectedbyflowcytometry.Witheachcelldivision,daughtercellsinheritroughlyhalfofthefluorescentlabel,allowingthenumberofcelldivisionsthatoccurafterlabelingtobedetectedbytheappearanceofsuccessivelydimmerfluorescentpeaksonaflowcytometryhistogramcomparedtocellsanalyzedimmediatelyafterstaining(Figs.1and2). Cellproliferationdyescanbeusedtotrackcelldivisionsinvivoorinvitro.ThestainingalsocanwithstandfixationandpermeABIlizationforsubsequentimmunostaining.
KitComponents:
Eachkitcontains10lyophilizeddyevials,anhydrousDMSOforpreparingstocksolutions,andalabelingprotocol.Onekitcanbeusedtolabelatleast1.8×108cells;thefinalnumberofassaysthatcanbeperformedperkitdependsonthedyeconcentrationused(seeproductprotocolunderdownloads).
ViaFluorkitscomein3dyeoptions:
- ViaFluor™CFSE(alsoknownasCFDA-SE)canbedetectedintheFITCchannel.
- ViaFluor™405-SEcanbedetectedinthePacificBluechannel.
- ViaFluor™568-SEcanbedetectedintheTexasRedorPE-TexasRedchannels.
ViaFluor™CFSEcanbeusedasacytoplasmicstaininyeast.HoweverViaFluor™405-SEandViaFluor™568-SEdonotstainwellinyeast.
Carboxyfluoresceindiacetate,succinimidylester(CFDA-SEorCFSE)isalsoavailableseparately.
References
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