Help! non-denaturing PAGE methods
dildo at utexas.edu
Thu Oct 29 14:50:59 EST 1998
I am trying to separate 600-700kD protein complexes from brain homogenate
on a non-denaturing gel. However it has not been successful. I reallyneeds
some suggestions from you.
I use 4-12% gel including 0.1% chaps instead of SDS. The running buffer is
Tris, glycine, and 0.1% chaps, and it is circulated during running PAGE.
The samples (25-100 micro g on each lane of a regular size gel) are run
24-48 hrs at constant volt at 40, or constant current at 20mA, then
stained by Coomassie blue. Every solubulized samples (by SDS, RIPA, chaps)
were stuck on top of the stacking gel, did not get into the gel. However
the marker went through. I do not know what do to next.
If you have some suggestions, I really appreciate.
Please respond e-mail to yumiko at mail utexas.edu
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