Modified RIPA Buffer Supplemented with Nucleases
| Reagent | Final concentration |
|---|---|
| Tris-HCl (pH 7.4) | 50 mm |
| NaCl | 100 mm |
| NP-40 | 1% |
| Sodium deoxycholate (DOC) | 1% |
| Sarkosyl | 0.1% |
| Dithiothreitol (DTT) | 5 mm |
| MgCl2 | 5 mm |
| Glycerol | 3% |
| Benzonase (e.g., Sigma-Aldrich #######) | 0.5 µL/mL |
| RNase A | 0.2 mg/mL |
| Complete protease inhibitors with EDTA (Roche) | 1× |
| Phenylmethylsulfonyl fluoride (PMSF) | 5 mm |
| Use buffer immediately, or prepare and keep it without enzymes and DTT at 4°C for several weeks. Add the enzymes and DTT immediately before use. | |
- © 2017 Cold Spring Harbor Laboratory Press
