Analysis of apoptosis using Xenopus egg extracts.

In the presence of a subcellular fraction enriched for mitochondria, after prolonged incubation the Xenopus egg extract can mimic biochemical aspects of apoptosis such as caspase activation, and DNA fragmentation. This unit describes preparation of an apoptotic extract from a crude interphase extract and an extract fractionated into latent and execution phases. An apoptotic extract can also be reconstituted from a fractionated interphase extract and purified mitochondria. Protocols are also included for monitoring apoptotic progression in the extract either by following activation of apoptotic proteases (caspases) or by assessing translocation of cytochrome c from the mitochondria to the cytosol.