Assaying cytochrome C translocation during apoptosis

Methods Mol Biol. 2004:284:307-13. doi: 10.1385/1-59259-816-1:307.

Abstract

Translocation of proteins from the mitochondrial intermembrane space to the cytoplasm is a critical event during apoptosis. There are several methods for assaying this event cited in the literature. In this chapter, we highlight separation of cytosolic and mitochondrial fractions of cultured cells using digitonin as the method for measuring cytochrome c release that, in our hands has been the simplest and most reproducible.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis*
  • Blotting, Western
  • Cytochromes c / analysis
  • Cytochromes c / metabolism*
  • Cytosol / enzymology
  • Cytosol / metabolism
  • Flow Cytometry
  • HeLa Cells
  • Humans
  • Intracellular Membranes / drug effects
  • Intracellular Membranes / enzymology
  • Intracellular Membranes / metabolism
  • Mitochondria / enzymology
  • Mitochondria / metabolism*
  • Subcellular Fractions / metabolism

Substances

  • Cytochromes c