Katherine Walstrom wrote:
> I have some dilute protein samples that I need to concentrate and run on
> an SDS-PAGE gel. I can't find a detailed protocol for precipitating
> proteins, although what I've read suggests that ethanol works well. I
> want to precipitate all the proteins, not just selectively precipitate
> some of them. Does anyone have a general protocol for this?
>> Thanks much.
>> Katherine M. Walstrom, PhD
> Division of Natural Sciences
> New College of USF
> 5700 N. Tamiami Trail
> Sarasota, FL 34243
>kwalstro at sar.usf.edu
What are you reading? There are several methods--3 vol of acetone works. I
like the methanol:chloroform method (Wessel & Flugge, Anal Biochem
138:141-3). You can also try using deoxycholate (Gates, Anal Biochem
196:290-). Most of these need to be resolubilized in the presence of a
small amount of SDS, which means that 1x SDS-PAGE buffer works swell.
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
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