precipitating proteins w/ ethanol

Kresten Lindorff Larsen kresten at my-dejanews.com
Mon Mar 8 10:52:29 EST 1999


As others, I always use TCA:
1. add 1/10 vol of
  100%   TCA, that is 100 mg pr. 100 ml final solution
    0.2% Sodium Deoxycholate, 0.2 mg pr. total 100 ml

The deoxycholate precipitates when diluted and helps to bring down the protein
precipitate

2. spin like hell in a tabletop microfuge at 4 degrees celcius. No need for
ice. I always spin about 20 min., but perhaps shorter times will also work.

3. remove supernatant and redissolve in an appropriate amount of 2x SDS-PAGE
loading buffer

4. if the sample turns yellow add just enough base, eg. 1M Tris or NaOH to
make it blue again

Good luck
Kresten


The address kresten at my-dejanews.com is for spambots only. Please mail me
at LysLeuLeu at crc.dk , transforming the pre at -part into my initials.
Kresten Lindorff Larsen, Dept. Yeast Genetics
Carlsberg Laboratory, Denmark

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