Western blot on Coomassie stained SDS-PAGE gel?
bjm10 at cornell.edu
Tue Feb 19 22:45:53 EST 2002
qqin at ccrc.uga.edu (Qiang Qin) wrote in news:3C6C8B26.75487142 at ccrc.uga.edu:
> Has anybody tried to eletroblot Coomassie stained gel directly to
> membrane then do Western? Will that work? Will the protein transfer
> well? And will the 1st antibody still react with the protein? Thanks.
As others have pointed out, this depends on the protein. However, there
are ways to force proteins to resolublize. I've never done it myself (and
don't have a proper protocol), but I recall that one method involved
soaking the gel in 6M urea for a while, then equilibrating in transfer
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