Caspases: determination of their activities in apoptotic cells.

摘要: Caspases, a family of cysteine proteases, were identified as major regulators apoptotic cell death. These enzymes are involved not only in initiation but also an execution phase apoptosis by cleaving more than 400 substrates. This cleavage mediates majority the typical biochemical and morphological changes cells, such shrinkage, chromatin condensation, DNA fragmentation, plasma membrane blebbing. In addition to their role death, caspases fulfill various nonapoptotic functions living cells. Thus, detection caspase activation/activity is essential for understanding different biological processes. It can be used marker induced diverse stimuli. chapter describes set methods available characterization and/or quantification activation, including immunoblotting, synthetic substrates, affinity labeling, flow cytometry microscopic techniques. addition, several discussing attempts vivo analyzing activity included. Advantages disadvantages each method compared.