NP-40 lysis buffer recipe
A milder alternative to RIPA for cytoplasmic and membrane proteins: 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP-40.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| 1 M Tris-HCl (pH 8.0) | 2.5 mL | 50 mM |
| 5 M NaCl | 1.5 mL | 150 mM |
| NP-40 (or IGEPAL CA-630) | 500 µL | 1% |
| Water (distilled) | to final volume |
Steps
- Combine the Tris and NaCl stocks and NP-40 with water and mix gently.
- Bring to the final volume with water.
- Store at 4 °C.
- Add protease inhibitors (and phosphatase inhibitors if needed) just before use.
Good to know
- Nuclear proteins are poorly extracted — use RIPA for whole-cell lysates.
FAQ
Is it suitable for Co-IP?
Yes — without SDS or deoxycholate, protein interactions are better preserved.
This is a widely used standard composition. Amounts depend on the hydrate form and purity of your reagents — check the formula weight (FW) on the bottle and follow your institution's safety rules and the manufacturers' instructions.