NucView™488Caspase-3substrateisanovelcellmembrane-permeablefluorogeniccaspasesubstratedesignedfordetectingcaspase-3activitywithinlivecellsinrealtime.VisitourProductVideopage toseeavideoofNucView™488inaction.

Therateofapoptosistypicallyvariesfromcelltocellevenwithinthesamepopulation.Asaresult,variousapoptoticeventsorMarkersaccompanyingtheapoptoticprocessalsooccurdifferentlyamongcells.Thus,itisimportanttobeabletodetecttheseapoptoticeventsonanindividualcellbasis.TrADItionally,caspaseactivityhasbeendetectedeitherusingamembrane-impermeablefluorogenicenzymesubstratesuchasDEVD-R110,orafluorescently-labeledinhibitorsuchasaFLICAreagent.Intheformercase,celllysisisrequired,thusprecludingthedetectionofcaspaseactivityinlivecells.Inaddition,suchcaspaseassaysmeasureonlytheaveragecaspaseactivityofahighlyheterogeneouscellpopulationatagiventime.Inthelattercase,althoughaFLICAreagentcanenterlivecellstodetectcaspaseactivity,onlytheinitialfluorescentsignalfollowingtheapplicationofthereagentcantrulyreflecttheenzymeactivityorthestateoftheapoptoticcellsbecauseanydetectedsignalaftertheinitial“snapshot”willneedtoconsiderthepotentialinterferenceoftheinhibitortotheenzymeandtheapoptoticcellitself.

Unlikeconventionalcaspaseassays,NucView™488Caspase-3substratedetectscaspase-3activitywithinindividualintactcellswithoutinhibitingcaspase-3activity.ThesubstrateconsistsofafluorogenicDNAdyeandaDEVDsubstratemoietyspecificforcaspase-3.Thesubstrate,whichisbothnon-fluorescentandnonfunctionalasaDNAdye,rapidlycrossescellmembranestoenterthecytoplasm,whereitiscleavedbycaspase-3toformahigh-affinityDNAdyethatstainsthenucleusbrightgreen.Thus,theNucView™488caspase-3substrateisbi-functional,allowingdetectionofintracellularcaspase-3activityandvisualizationofchangesinnuclearmorphologyduringapoptosis.Thefluorescentstainingproducedinresponsetocaspase-3activityiscompatIBLewithsubsequentfixationandpermeABIlizationforimmunostaining.

ThekitcontainsNucView™488Caspase-3substrateandacaspase-3inhibitorAc-DEVD-CHOsufficientfor100flowcytometryormicroplateassays(200uLassayvolume).Thetrialsizekitcontainssufficientreagentsfor25assays(200uLassayvolume).Thenumberoffluorescencemicroscopyassaysthatcanbeperformedwiththekitmayvarybasedonthesizeofculturevesselandstainingvolumeused.Thekitprovidesaconvenienttoolforprofilingapoptoticcellpopulationbasedoncaspase-3activityusingeitherfluorescencemicroscopyorflowcytometry.

KitComponents:

• NucView™488Caspase-3substrate,0.2mMinDMSO
• Caspase-3inhibitorAc-DEVD-CHO,2mMinDMSO

Features:

• Realtimecaspase-3detectioninintactcells:Detectcaspase-3activitywithinindividualcellsinapopulation.
• Bi-functional:Allowsvisualizationofcaspase-3activityandapoptoticnuclearmorphologyatthesametime.
• Simple&Fast:15-minuteassaywithnowashrequired.
• Versatile:Fordetectionbyflowcytometry,fluorescencemicroscopy,orfluorescencemicroplatereaderusingfluoresceindetectionsettings.
• Fixable:Compatiblewithstandardfixationandpermeabilizationmethodsforsubsequentimmunofluorescencestaining.

SeeourfullselectionofNucView™substratesandkits.

NucView™enzymesubstratetechnologyiscoveredbyU.S.PatentNos.8,092,784and8,586,325.