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
4. if the sample turns yellow add just enough base, eg. 1M Tris or NaOH to
make it blue again
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
-----------== Posted via Deja News, The Discussion Network ==----------
http://www.dejanews.com/ Search, Read, Discuss, or Start Your Own