| Triton X-100 (Sigma-Aldrich T8787) |
52.5 µL |
0.5% |
| Tris-HCl (1 m, pH 7.5; ThermoFisher 15567-027)
|
105 µL |
10 mm |
| CaCl2 (1 m; Sigma-Aldrich C3306)
|
31.5 µL |
3 mm |
| Sucrose (Sigma-Aldrich S0369) |
882 mg |
250 mm |
| DTT (0.1 m; Sigma-Aldrich D9779)
|
10.5 µL |
0.1 mm |
| Complete Mini EDTA-free Proteinase Inhibitor Cocktail (Roche 11836170001) |
1.5 tablets |
1× |
| Phenylmethylsulfonyl fluoride (PMSF; 0.1 m in isopropanol; Fluka 93482)
|
105 µL |
1 mm |
| Prepare the solution fresh before use. Add protease inhibitors (Complete Mini EDTA-free Proteinase Inhibitor Cocktail and
PMSF) immediately before using the buffer.
|
