Cytochrome C western blot

Jayakumar R jakku71 at
Sat Dec 8 15:34:31 EST 2001

   I am having a strange problem. I am doing western blots to detect
Cytochrome C  of human mitochondrial total protein extracts.  the Ab is a
mouse monoclonal Ab (Zymed) against Cyt C sensitive to Human Cyt C, expected
to give a band at 15 kDa.  The Ab is senstive only to denatured Cyt C. I
always happen to get a major band at near ~56 kDa position and a very weak
band at 15 kDa.  I tried increasing the concentration of ME upto 15% (1X
conc.) and still no use.  I was wondering whether it is some sort of a
tetramer of CytC. Can anyone explain what is going on here?
  By the way, the gel is a 15% SDS-PAGE gel run on a mini protean II
aparatus. Any help will be gratefully appreciated.

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