蛋白質沉澱

TCA/ ACETONE Protein precipitation
Solution A: 10 % TCA, 0.07 %2-mecraptoethanol (2-MET) / in 100 % acetone
Solution B: 0.07 % 2-MET, 1 mM PMSF/ in 80 % acetone
Store at -20～80 oC

1. Sample weight: Solution A = 1:10, store at -20 oC for 45 min (no more than 1 hr)
2. Centrifugal,12,000 rpm, 15 min,  4 oC, remove the supernatant
3. Solution B wash x 3~5 times
4. 100 % Acetone wash x 1, As clean as possible
5. Air dry for 5 min
6. 100 uL rehydration buffer

Solution A: 10 % TCA, 0.07 %2-mecraptoethanol (2-MET) / in 100 % acetone
Solution B: 0.07 % 2-MET, 1 mM PMSF/ in 80 % acetone
Store at -20～80 oC

Rehydration buffer
8M urea. 2 % CHAPS, 0.5% IPG buffer, 0.002% Bromophenol blue
When need to use, add 20 mM (0.0031 g/mL DTT)