Overview | TheGuavaCaspaseAssaySuiteoffluorescent-basedassaysforCaspase 3/7,8,9andmulti-caspaseemploythesameprotocolmakingiteasier toperformmultiplecaspaseassaysinparallel.Further,Guavacaspase reagentsaremembranepermeableallowingonetomeasureintracellular levelsoftheenzymewithoutusingharshlysismethods.TheGuava CaspaseAssaysemployaFLICA(FluorescentLabeledInhibitorof Caspase)reagentthatspecificallyidentifiesactivecaspases.EachFLICAis comprisedofthreesubunits: –anactivesitebindingpeptidetoconferspecificity(DEVDforcaspase3/7, LETDforcaspase8,LEHDforcaspase9,andVADformulti-caspase) –acovalentcrosslinkingagent(FMK) –agreen(FAM)ororange-redfluorescentprobe(SR):SRisexcitedbyblue orgreenlasers(PCASystemsorEasyCyteSystems),FAMbybluelasers only(EasyCyteSystems) TheGuavaCaspaseAssaysalsoincludethenuclearDNAstain,7-AAD,to simultaneouslyevaluatemembraneintegrityandcellviABIlity. |