Analysing caspase activation and caspase activity in apoptotic cells.
Apoptotic cell death is characterised by various morphological and biochemical changes. Cysteine proteases of the caspase family play key roles in the execution of apoptosis and in the maturation of proinflammatory cytokines. During apoptosis signalling, caspase precursors undergo rapid proteolytic processing and activation. Activated caspases then function to cleave various vital cellular proteins, resulting in the death of the cell. Thus, the measurement of caspase activation and caspase activity provides a quick and convenient method to assess apoptosis. This chapter outlines various commonly used assays for measuring caspase activity and detecting active caspases in cultured cells or tissue extracts.
Kumar
Sharad
S
Hanson Institute, Adelaide, Australia.
Dorstyn
Loretta
L
D000818:Animals; D017209:Apoptosis; D001671:Biochemistry / Q000379:methods*; D020169:Caspases / Q000032:analysis* / Q000378:metabolism*; D002478:Cells, Cultured; D004586:Electrophoresis / Q000379:methods; D004789:Enzyme Activation; D006801:Humans; D015151:Immunoblotting / Q000379:methods
D020169:Caspases