RIPA lysis buffer recipe
The most common lysis buffer for extracting protein from cells and tissue, including nuclear and membrane proteins. 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS.
🧮 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% |
| Sodium deoxycholate | 250 mg | 0.5% |
| 10% SDS | 500 µL | 0.1% |
| Water (distilled) | to final volume |
Steps
- Add the Tris and NaCl stocks to about 70% of the final volume of water.
- Dissolve sodium deoxycholate, then add NP-40 and 10% SDS (avoid foaming).
- Bring to volume and store at 4 °C.
- Add protease inhibitors (and phosphatase inhibitors if needed) just before use.
Good to know
- Sodium deoxycholate is light-sensitive — store in a dark bottle.
- RIPA lysates are compatible with BCA but not with Bradford (detergents interfere).
FAQ
Can I add protease inhibitors in advance?
Many inhibitors lose activity in solution within hours to days — add them to the aliquot you use.
Is RIPA suitable for Co-IP?
RIPA is harsh and can break weak interactions; use a milder NP-40 or Triton-only buffer for Co-IP.
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.