Multiple antibodies in Western

Ian A. York iayork at panix.com
Mon Dec 2 12:14:26 EST 2002


In article <a07dedf1.0212020851.656b2bdf at posting.google.com>,
Jason Wong <7jw6 at qlink.queensu.ca> wrote:
>I'm probing for two presynaptic markers in guinea pig hippocampus,
>SNAP-25 and synaptophysin.  I've purchased mouse monoclonal antibodies
>from Stressgen and Dako respectively and have had excellent results
>after a couple trial gels.  I'm currently trying to get the whole
>stripping/reprobing thing working, but I had a question: can I
>incubate my blots with two different primary antibodies at the same
>time?  My bands are separated quite well (23 kDa and 38 kDa
>respectively) using a 12% Tris-HCL gel.

Sure, no problem.  it might help to tweak your antibody concentrations so 
the bands are of similar intensities, but if you're getting good single 
westerns you should also get good doubles or triples.

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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