Tris-EDTA pH 9.0 antigen retrieval buffer recipe
An alkaline heat-induced epitope retrieval buffer for formalin-fixed paraffin sections: 10 mM Tris, 1 mM EDTA, 0.05% Tween-20, pH 9.0.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Tris base | 1.21 g | 10 mM |
| 0.5 M EDTA (pH 8.0) | 2 mL | 1 mM |
| Tween-20 | 500 µL | 0.05% |
| Water (distilled) | to final volume |
Steps
- Dissolve in about 80% of the final volume of water.
- Adjust to pH 9.0 and add the Tween-20.
- Bring to the final volume with water.
- Store at room temperature for 3 months or at 4 °C.
- Heat sections at 95–100 °C for 20 min and cool in the buffer for 20 min.
Good to know
- Stronger than citrate pH 6.0 and often gives more signal, but sections detach more easily.
FAQ
Citrate or Tris-EDTA?
Follow the antibody datasheet first; if nothing is stated, test both side by side.
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.