Citrate buffer pH 6.0 (antigen retrieval) recipe
Used for heat-induced epitope retrieval (HIER) to unmask formalin-fixed antigens in paraffin IHC. 10 mM sodium citrate, 0.05% Tween-20, pH 6.0.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Trisodium citrate dihydrate | 2.94 g | 10 mM |
| Tween-20 | 500 µL | 0.05% |
| Water (distilled) | to final volume |
Steps
- Dissolve sodium citrate in about 900 mL of water.
- Adjust to pH 6.0 with 1 N HCl.
- Add Tween-20 and make up to 1 L.
- Store 3 months at room temperature, longer at 4 °C.
Good to know
- Some antigens retrieve better in Tris-EDTA pH 9.0 — follow the antibody datasheet.
FAQ
How long should I heat it?
Typically 10–20 min at 95–100 °C, then 20 min cooling at room temperature; microwaves, pressure cookers and water baths are all used.
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.