UNIT 11.12 Analysis of Apoptosis Using Xenopus Egg Extracts
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cell Biology
How to Cite
Kornbluth, S. and Evans, E. K. 2001. Analysis of Apoptosis Using Xenopus Egg Extracts. Current Protocols in Cell Biology. 9:11.12:11.12.1–11.12.8.
- Published Online: 1 MAY 2001
- Published Print: JAN 2001
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.