SDS-PAGE gel-shift to look at protein phosphorylation

John Pawlowski johnp at qadas.com
Tue Jan 13 12:02:48 EST 1998

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

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