Assaying cytochrome C translocation during apoptosis.
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.
Waterhouse
Nigel J
NJ
Cancer Immunology, Research Division, Peter MacCallum Cancer Center, Melbourne, Australia.
Steel
Rohan
R
Kluck
Ruth
R
Trapani
Joseph A
JA
D017209:Apoptosis; D015153:Blotting, Western; D045304:Cytochromes c / Q000032:analysis / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism
D045304:Cytochromes c