Description

Intheintrinsicapoptosispathway,mitochondrialpermeABIlitytransitionisanimportanteventwhereintheelectRochemicalgrADIent(referredtoasdelta-psiorΔΨm)acrossthemitochondrialmembranecollapses.Thiscollapseoccursthroughtheformationofchannelsorporesintheoutermitochondrialmembrane,whichinvolvesBaxinsertionandoligomerization,followedbythereleaseofCytochromeCintothecytoplasm.

TheDePsipher™Kitusesauniquecationicdye(5,5’6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolyl-carbocyanineiodide)toindicatethelossofΔΨm.Thedyereadilyenterscellsandfluorescesbrightlyredinitsmultimericformwithinhealthymitochondria.Inapoptoticcells,themitochondrialmembranepotentialcollapses,andtheDePsipher™reagentcannotaccumulatewithinthemitochondria.Inthesecells,DePsipher™returnstoitsgreenfluorescentmonomericform.Apoptoticcells,showingprimarilygreenfluorescence,arethuseasilydifferentiatedfromhealthycellswhichshowredfluorescence.Theaggregateredformhasabsorption/emissionmaximaof585/590nm,andthegreenmonomericformhasabsorption/emissionmaximaof510/527nm.Bothapoptoticandhealthycellscanbevisualizedsimultaneouslybyepifluorescencemicroscopyusingawideband-passfilter.

TheDePsipher™reagentiseasytouse.SimplyresUSPendthereagentinreactionbufferorculturemedia(withorwithoutthestabilizersolution),addtoyourcells,incubatefor15to20minutes,washandanalyzebyflowcytometryormicroscopy.Visualizationbymicroscopyallowsarapidinspectionandqualificationofapoptosis.Flowcytometricanalysisallowseasyquantitationofcelldeathasevidencedbymitochondrialpotentialbreakdown.

FEATURES:

• Simple.JustaddDePsipher™reagenttomediaorreactionbuffer.
• UniqueStabilizerSolutionimprovesresults.
• Fast.Takesonly20minutes.
• FlexIBLe.Viewcellsbyepifluorescenceorconfocalmicroscopy,oranalyzecellsbyflowcytometry.

APPLICATIONS:

• Flowcytometry
• Epifluorescencemicroscopy
• Confocalmicroscopy