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
| Catalog#6300-100-Kincludes: | ||
| CatalogNumber | Description | Qty |
| 6300-100-01 | DePsipher™ | 1 |
| 6300-100-02 | 10XReactionBuffer | 1 |
| 6300-100-03 | StabilizerSolution | 1 |
