Blotting onto PVDF

Dr E. Buxbaum EB15 at le.ac.uk
Mon Aug 3 17:09:56 EST 1998


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).



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