[Protein-analysis] Western blot on Coomassie stained SDS-PAGE gel?

kaydamov via proteins%40net.bio.net (by kaydamov At ipk-gatersleben.de)
Tue Nov 28 08:38:28 EST 2006


Dear Andreas Savelsbergh,

I was founding in an old forum, that you routinely use Coomassie stained
gels directly for electroblotting and subsequent antibody detection. I also
used Coomassie stained gels but until now only for immunoblotting on
PVDF-membranes. The Coomassie stain disappeared after soaking the
PVDF-membranes in 100% methanol prior blocking and antibody steps.
However, now I'm using nitrocellulose membranes where the methanol-step is
omitted. Without a methanol-step it seems that the Coomassie stain will
persist on the membrane also during the first blocking step and all the
following incubation steps. Does the Coomassie stain interfere with the
antibody binding or the chemoluminesent detection (I'm using ECL detection
system)? And if yes, is there a possibility to remove the Coomassie stain
from the NC-membrane before immunoblotting? Or is it only possible to use
Coomassie stained gels directly for immunoblotting only together with
PVDF-membranes?



I would be very grateful for some helpful informations.



Sincerely yours



Catrin Kaydamov



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