ECL Western Blotting, need help

Ian A. York iayork at panix.com
Thu Aug 7 08:13:28 EST 1997


On 31 Jul 1997 00:19:25 GMT, yankiwski at aol.com (Yankiwski) wrote:
>
>>I've been trying to detect a protein transferred to Immobilon membranes,
>>and have had poor results.  On one attempt I got lots of bands with high

Are you positive that your proteins are binding well to the PVDF?  

I was using PVDF with no problems for quite a while; got a new batch in
and suddenly my westerns stopped working.  After fooling around a bit I
tried backing the PVDF with nitrocellulose and found that perhaps 90% of
the protein was zipping right through the PVDF.  Switching back to
nitrocellulose solved my problems.

Of course, that won't account for your high background, so I guess I'm
just babbling again.

Ian
-- 
      Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
      "-but as he was a York, I am rather inclined to suppose him a
       very respectable Man." -Jane Austen, The History of England



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