High-salt lysis buffer composition for HCF-1 protein immunoprecipitation
| Reagent | Stock | Final concentration (mM or μM) | Amount (μL or mL) |
|---|---|---|---|
| Tris pH 7.5 | 1 M | 50 Mm | 5 mL |
| NaCl | 4 M | 300 mM | 7.5 mL |
| Triton X-100 | 10% | 0.5% | 5 mL |
| EDTA | 500 mM | 5 mM | 1 mL |
| DTT | 500 mM | 1 mM | 200 μL |
| PMSF | 100 mM | 1 mM | 1 mL |
| Anti-protease | 500 mM | 1:100 | 1 mL |
| NaF | 500 mM | 50 mM | 10 mL |
| βGP | 1 M | 10 mM | 1 mL |
| Na3VO4 | 200 mM | 1 mM | 500 μL |
| Sodium Butyrate | 200 mM | 10 mM | 1 mL |
| PUGNAc | 2 mM | 10 μM | 500 μL |
| ddH2O | n/a | n/a | 66.3 mL |
| Total | n/a | n/a | 100 mL |
Store at 4°C. The minimal buffer is stable for a year.