IUBio

precipitating proteins w/ ethanol

Rich Dudley rdudley+ at pitt.edu
Fri Mar 5 09:10:22 EST 1999


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.

Good luck!

rich


--
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
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