ACK red blood cell lysis buffer recipe
Lyses red blood cells in blood or spleen preparations: 150 mM NH₄Cl, 10 mM KHCO₃, 0.1 mM EDTA.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| NH₄Cl | 4.01 g | 150 mM |
| KHCO₃ | 500 mg | 10 mM |
| Na₂EDTA·2H₂O | 18.6 mg | 0.1 mM |
| Water (distilled) | to final volume |
Steps
- Dissolve in about 80% of the final volume of water.
- Adjust to pH 7.2–7.4 (1 N HCl).
- Bring to the final volume with water.
- Filter-sterilise through 0.22 µm. Store at room temperature.
Good to know
- Add 1–5 mL to the pellet, wait 3–5 min at room temperature, then stop with 10 volumes of PBS.
- Longer exposure damages leukocytes too.
FAQ
What about hypotonic water lysis?
Brief water lysis followed by 10X PBS works, but timing is tricky, so ACK is more common.
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.