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).
