First I'd like to thank everyone for their responses! The gel that she
ran to today was much improved. We are still not exactly sure of the
problem. Perhaps it was Triton or residual salts. The way we got rid
of the problem for anyone with the same problem, was to EtOH ppt the
proteins for ~30 min at -20 C. Air-dried O/N. The pellet was
resuspended in 0.0625 M Tris. Run through a microconcentrator (low MW
cutoff). Added tris (wash) to max volume of microconcentrator three
times. Spun down to yield a final volume of ~50 ul. It's a hell of a
lot of work, but the band diffusion was much, much less. We also ran
the gel slower. But once again, I would like to thank all of you who
offerred your advice. I'd hope I could do the same for you some day.
Thanks.
Sincerely, Joe Stains Penn State University
