| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 490/515 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.Foreachsample,preparecellsin0.5mLwarmmediumorbufferofyourchoiceatadensityof5×105to1×106cells/mL.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforapoptosisinduction.
2.Treatcellswithtestcompoundsforadesiredperiodoftimetoinduceapoptosis,andcreatepositiveandnegativecontrols.
3.Add1μLof500XTF2-VAD-FMK(ComponentA)intothetreatedcells(fromStep2),andincubatethecellsina37°C,5%CO2incubatorfor1-4hours.
Note1:Foradherentcells,gentlyliftthecellswith0.5mMEDTAtokeepthecellsintact,andwashthecellsoncewithserum-containingmediapriortoincubationwithTF2-VAD-FMK.
Note2:Theappropriateincubationtimedependsontheindividualcelltypeandthecellconcentrationused.Optimizetheincubationtimeforeachexperiment.
4.Washandspinthecellstwice.ResUSPendthecellsin0.5mLofassaybufferorgrowthmedium.
Note:TF2-VAD-FMKisfluorescent,thusitisimportanttowashoutanyunboundreagenttoremovethebackground.
5.Ifdesired,labelthecellswithaDNAstain(suchaspropidiumiodideor7-AADfordeadcells).
6.Ifdesired,fixcells.
7.MonitorthefluorescenceintensitywithaflowcytometerusingtheFL1channel(Ex/Em=490/525nm).Gateonthecellsofinterest,excludingdebris.
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