Blotting onto PVDF
srman at gengenp.rug.ac.be
Wed Aug 5 03:12:31 EST 1998
Dr E. Buxbaum wrote:
> Karen spink wrote:
> > Has anyone had any problems Western blotting onto PVDF in CAPS buffer? I am
> > trying to get a sequence on my protein and it appears that the efficiency
> > of transfer is letting me down. Could it be the buffer, (10mM CAPS, 10%
> > methanol, pH 11) ? I have had no problems blotting onto nitrocellulose
> > using tris-glycine buffers.
> It may be the high detergent concentration which is causing problems. If
> you want to avoid the glycine, why not use Dunn's buffer (published in
> Anal. Biochem. a couple of years back, NaHCO3/Na2CO3, which can be
> supplemented with methanol or SDS as needed). I use it routinely
> (because I found the transfer efficiency better than with tris-gly, and
> also because it is a lot cheaper).
i read about your problems with caps buffer, we too here in the lab used
this buffer , but i also tried to blot on pvdf with tris glycine buffer
and it works good too.
try it and let me know.
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