| Tris-HCl (1 m, pH 8.0)
|
10 mm |
| Sucrose (1 m)
|
0.3 m |
| MgCl2 (1 m)
|
5 mm |
| KCl (1 m)
|
25 mm |
| NaF (500 mm)
|
10 mm |
| β-Glycerophosphate (1 m)
|
5 mm |
| Sodium pyrophosphate (100 mm)
|
5 mm |
| Spermine (500 mm)
|
0.5 mm |
| Spermidine (500 mm)
|
0.5 mm |
| Pefabloc SC PLUS (Roche 11873601001) (20 mg/mL) |
0.1 mg/mL |
| Dithiothreitol (DTT) (1 m)
|
2 mm |
| Protease inhibitor tablet (cOmplete, EDTA-free) (Roche 4693132001) |
1 tablet for 50 mL |
| Combine Tris-HCl, sucrose, MgCl2, KCl, NaF, β-glycerophosphate, sodium pyrophosphate, spermine, and spermidine. Store the buffer at 4°C. Immediately before
use, add Pefabloc SC, DTT, and protease inhibitor.
|
