help!how to detect cytochrome C releasing from motocondria!

[Paul S. Brookes.]

Lyse (gently) the cells and centrifuge.  Then western blot for cyt-c in the 
mitochondrial pellet and the cytosol/supernatant.  Pharmingen's mouse 
monoclonal Ab works well in our hands.  See FASEB J. vol 13, pp2031-2036 
(1999) for  a recent example of this.

Of course with any such method you have to make sure any cyt-c you see in 
the supernatant is actually released and not due to non-pelleted 
mitochondria.  Enzyme assays like citrate synthase are simple enough to 
accomplish this.

Regards
PSB


_________________________________________
Dr. Paul S. Brookes.            (brookes at uab.edu)
UAB Department of Pathology,   G004 Volker Hall
1670 University Blvd., Birmingham AL 35294 USA
Tel (001) 205 934 1915     Fax (001) 205 934 1775
http://peir.path.uab.edu/brookes

The quality of e-mails can go down as well as up
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