In article <ctcuu9$6iv$[Only registered users see links. ].uni-hamburg.de>, "news.rrz.uni-hamburg.de" <[Only registered users see links. ]> wrote:
Tris-Glycine: no good reason. I think the correct explanation is historical:
Towbin took 2X diluted SDS-PAGE running buffer without SDS and it
worked. As long as the stuff conducts but does not heat much (e.g.,
low moving ions), pH is high enough and the buffer is good enough,
anything should work. There are certainly other alternatives, like
(bi)carbonate or CAPS.
Methanol: To decrease polarity of the solution and to keep proteins
denatured. Both factors promote strong binding of ptoteins to the
I know Amersham have a manual inclosed in the box with the
western membranes that give out the reference to that question. If I
remember correctly, PDVF membranes require methanol in their transfer
buffer but not nitrocellulose ones. Check in your suppliers box, see
if they have that manual.