precipitating proteins w/ ethanol
i.mcfarlane at icrf.icnet.uk
Thu Mar 11 10:08:24 EST 1999
Unfortunately my protein does not precipitate in TCA at all.
Thanks for your reply tho.
> 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
> 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
> 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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