acetone precipitation of samples (for SDS PAGEs)

Sai siyer at bmg.bhs.uab.edu
Fri Nov 15 17:23:27 EST 1996


hey all, 
            i've posted this before, but i figured i'd try again to see if
there were any other ideas.  i've been doing acetone precipitation to
remove salts from my samples prior to loading on sds gels.  the basic
protocol is add 8 vol. of acetone to 1 vol. of protein sample --->
precipitate at -20 degrees for 4-6 hours ---> spin down for 20 min at
14,000g (max speed on centrifuge) at 4 degrees to pellet out the
precipitated protein ----> decant the supernatant and immediately
solubilize the pellet in sds loading buffer.  the problem is see is that i
still get streaks in my gels.  theoretically, acetone is an uncharged
organic solvent and so should not create any distortions in the gel cuz
its not charged.  so if there were any residual acetone left, that still
shouldnt create the streaks...yet i still see streaks.  anybody have any
ideas on this?  or other methods to remove salts from samples (i.e tca
precipitations etc) would also be welcome...thx...cheers....

-- 
Sai Iyer
siyer at bmg.bhs.uab.edu
graduate student; dept of biochemistry and molecular genetics
university of alabama at birmingham



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