Hi all,
I'm currently looking at the phosphorylation state of a protein by
SDS_PAGE gel shift analysis. The protein is about 46 kDa and a noticible
shift is seen using 8% polyacrylamide, 1.5 mm gel, approx 15 cm long.
Unfortunately the sample is very near the bottom of the gel. I would like
to increase the resolution of the gel-shift. Using 6 or 7% gels does not
allow good separation of the protein from the dye front. Someone has
suggested to me that I try running the sample longer using an 8% gel in a
DNA sequencing gel apparatus (0.2 or 0.4 mm thick). Any thoughts on
whether or not this approach would work or have any other ideas? ie.
playing with the pH of the resolving gel, concentrations of cross-
linkers? Any advice will be greatly appreciated. Thanks in advance.
John Pawlowski
johnp at qadas.com
