NucView™488andRedDot™2Apoptosis/NecrosisAssayKitcontainsNucView™488Caspase-3Substratefordetectionofcaspase-3/7activity.Thefar-reddead-cellstainRedDot™2isincludedforstainingnecroticandlateapoptoticcellsthathavecompromisedplasmamembraneintegrity.Thiskitprovidesaconvenienttoolforprofilingapoptoticandnecroticcellpopulationsbyfluorescencemicroscopyorflowcytometry.
Incontrasttootherfluorogeniccaspasesubstratesorfluorescentcaspaseinhibitorbased(FLICA)assays,NucView™488Caspase-3Substratecanbeusedtodetectcaspase-3/7activitywithinindividualintactcellswithoutinhibitingapoptosisprogression.ThesubstrateconsistsofafluorogenicDNAdyecoupledtothecaspase-3/7DEVDrecognitionsequence.Thesubstrate,whichisinitiallynon-fluorescent,penetratestheplasmamembraneandentersthecytoplasm.Inapoptoticcells,caspase-3/7cleavesthesubstrate,releasingthehigh-affinityDNAdye,whichmigratestothecellnucleusandstainsDNAwithbrightgreenfluorescence.Thus,NucView™488Caspase-3Substrateallowsdetectioncaspase-3/7activityandvisualizationofmorphologicalchangesinthenucleusduringapoptosis.
RedDot™2isacellmembrane-impermeable,far-reddyewithhighselectivityformembranecompromisedordeadcells.RedDot™2hasfar-redemissionat695nm,well-separatedfromthegreenfluorescenceofNucView™488. TheexcitationmaximumofRedDot™2is665nm,butitcanbeefficientlyexcitedbywavelengthsfrom488to647nm,andthereforecanbeusedwiththe488nmflowcytometrylaserline.
Note:whileNucView™488stainingisformaldehyde-fixableandcompatIBLewithsubsequentimmunostaining,fixationafterstainingwithRedDot™2isnotrecommendedbecauseitcanresultinincreasedbackgroundstainingofhealthycells.
SeeourfullselectionofNucView™substratesandkits.
NucView™enzymesubstratetechnologyiscoveredbyU.S.patents.
References
Cen,etal.DEVD-NucView488:anovelclassofenzymesubstratesforreal-timedetectionofcaspase-3activityinlivecells.TheFASEBJournalpublishedonlineFebruary8,2008.
Downloadafulllistofreferences:ValidatedcelllinesforNucView488withreferences
