1 M HEPES (pH 7.4) recipe
A buffering agent for culture media and protein buffers, effective at pH 7.2–7.6 and less temperature-sensitive than Tris.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| HEPES (free acid) | 23.83 g | 1 M |
| 10 N NaOH | as needed | to pH 7.4 |
| Water (distilled) | to final volume |
Steps
- Dissolve HEPES in about 80% of the final volume.
- Adjust to pH 7.4 with 10 N NaOH.
- Bring to volume and filter through 0.22 µm.
- Store at 4 °C protected from light.
Good to know
- HEPES can generate hydrogen peroxide under light, so keep it in the dark.
FAQ
Can I autoclave HEPES?
Filter sterilisation is recommended.
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.