Blotting of Coomassie stained gels possible?
PETER.WILSON at BBSRC.AC.UK
Tue Sep 2 08:09:36 EST 1997
<Pine.WNT.3.95.970902115217.-101881G-100000 at eliza.mpimf-heidelberg.mpg.de>,
stolpe at MZF.MPIMF-HEIDELBERG.MPG.DE ("Andreas M. Stolpe") wrote:
> Just a short question: Is Western Blotting of Coomassie stained PAGE
> possible? Do you have some recipes or suggestions?
Western blotting of coomassie stained PAGE does not work at all well since
the proteins are fixed by the methanol/acetic acid mix for the stain. If
you want to blot a protein gel after staining it I would reccomend using
Imidazole-zinc. Biorad sell a kit but it is much cheaper to DIY from the
C.Fernandez-Patron, L.Castellanos-Sera and P.Rodriguez Biotechniques
(1992) vol12 no4 pp 564-573
It is more sensitive than coomassie and uses only two solutions. It only
takes 30 minitues to stain and 15 minutes to destain before blotting. The
only hiccup is that it is a reverse stain (clear bands on a white
background) so you need to photograph it against a black background.
Hope this helps.
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