InductionofApoptosis.........4MeasurementofApoptosis-InducedChangesatthePlasmaMembrane...4MeasurementofApoptosis-InducedChangesintheMitochondria......7MeasurementofApoptosis-InducedChangesintheCytoplasm.......10MeasurementofApoptosis-InducedChangesintheNucleus.........17MeasurementofCellProliferation...21APPENDIXTablesTableI:CharacterizationofAntibodiestoCaspases.......22TableII:ClassificationofBcl-2FamilyMembers.......24TableIII:InducersofApoptosis...25TableIV:CaspaseInhibitors.....28Protocols:I.InductionofApoptosis......30II.CaspaseAssays(ColorimetricandFluorometric)...30III.GeneralGuidelinesforUsingFluorometricSubstratestoMeasureCaspaseActivity...........33Tools&TipsforAnalyzingApoptosis:AKitSelectionGuideCalbiochem•ToolsandTipsforAnalyzingApoptosis:AKitSelectionGuide1Overthepastthreedecades,thestudyofapoptosis(pro-grammedcelldeath)hasgainedsignificantimportanceinhumandiseaseanditsclinicalmanagement.Failuretoregulateapoptosisisacommonfeatureinseveraldiseasesincludingautoimmunedisorders,neurodegen-erativediseases,cancer,andAIDS.Hence,itisimportanttounderstandtheapoptoticprocessesincellsinordertodevisetherapeuticmeanstointerveneandresetthebal-ancebetweencellsurvivalanddeath.Itisnowwellrecog-nizedthattherearetwomainpathwaysfortheinductionofapoptosis,theextrinsicorreceptor-mediatedpathway,andtheintrinsicormitochondrialpathway.Inductionofeitherpathwaycanresultintheactivationofcaspases,aclassofintracellularcysteineproteasesthatarerespon-sibleforthecleavageofavarietyofcellularsubstratesandthemorphologicalchangesattributedtoapoptosis.Otherlesswell-definedcaspaseactivationpathways,suchasautophagy,haverecentlybeendescribed,butwillnotbediscussedhere.Averylargebeneficiaryofapoptosisresearchisoncology,sincemostcancercellsexhibitdefectsintheirsuicidalmachinery.Bybetterunderstandingcaspaseactivationpathways,newtherapeuticagentsmaybedevelopedtoinducedeathincancerouscells.Ontheotherhand,phar-macologicinterferencewiththeinductionorcompletionofapoptosisholdspromiseforthetreatme...
