NucView™405Caspase-3Substrate,1mMinPBS,providesaconvenienttoolfordetectingapoptosisinintactcellsbasedoncaspase-3/7activityusingeitherconfocalmicroscopyorflowcytometry.
Incontrasttootherfluorogeniccaspasesubstratesorfluorescentcaspaseinhibitorbased(FLICA)assays,NucView™caspase-3substratescanbeusedtodetectcaspase-3/7activitywithinindividualintactcellswithoutinhibitingapoptosisprogression.NucView™substratesconsistofafluorogenicDNAdyecoupledtothecaspase-3/7DEVDrecognitionsequence.Thesubstrate,whichisinitiallynon-fluorescent,penetratestheplasmamembraneandentersthecytoplasm.Inapoptoticcells,caspase-3/7cleavesthesubstrate,releasingthehigh-affinityDNAdye,whichmigratestothecellnucleusandstainsDNAwithfluorescence.Thus,NucViewcaspase-3substratesarebifunctional,allowingdetectionofcaspase-3/7activityandvisualizationofmorphologicalchangesinthenucleusduringapoptosis.Thestainingisalsoformaldehyde-fixable.
NucView™405 dyeisexcitedbythe405nmlaserlinefordetectionbyconfocalmicroscopyintheDAPIchannel,orbyflowcytometryinthePacificBlue®channel.NucView™405isidealforcaspase-3detectioninmulti-colorapplicationsforresearcherswhowishtoreservethegreenfluorescencechannelforotherdetectionreagents.
NucView™405Caspase-3Substrateisofferedasa1mMsolutioninDMSO(catalogno.10405)orPBS(phosphate-bufferedsaline).ThesubstrateinPBSisformulatedforuseincellsthataresensitivetoDMSOtoxicity.Innon-DMSOsensitivecelltypes,addingDMSOduringthesubstrateincubationmayenhanceNucView™405staining.
Note:NucView™405signalmaybedifficulttoobserveusingepifluorescencemicroscopy.Confocalmicroscopyusing405nmlaserexcitationisrecommendedforimaging.BiotiumalsooffersgreenfluorescentNucView™488caspase-3substrateandkitsaswellasorangefluorescentNucView™530.
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