electrotransfert of LMW proteins

Ian A. York iayork at panix.com
Tue Apr 15 09:43:22 EST 1997


In article <5ittgk$7bk$1 at storm.worldnet.net>,
David Rouquie  <rouquie at ensam.inra.fr> wrote:
>
>Does anyone have experience with the electroblotting of low molecular
>weight proteins (< 20 kD)? 

I've done westerns on several proteins of 10 or 12 kDa and had no
problems.  If you're getting poor results, the most likely problem is that
the protein is going right through your blot.  I use the low molecular
wegiht transfer buffer recommended by Harlowe (in Antibodies: A Lab
Manual; this is the transfer buffer without SDS).  For the blot I've used
PVDF (my current favourite) and nitrocellulose; I believe that results may
be more variable with different batches of ntirocellulose, and I think you
might need to check pore size.  My transfer conditions and so forth are no
different than for medium sized (e.g. 50-100kDa) proteins.

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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