0.1 M glycine-HCl (pH 2.5) recipe
An acidic buffer for eluting antibodies from Protein A/G or antigen columns. Neutralise immediately with 1 M Tris pH 8.5.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Glycine | 3.755 g | 0.1 M |
| Concentrated HCl | as needed | to pH 2.5 |
| Water (distilled) | to final volume |
Steps
- Dissolve in about 80% of the final volume of water.
- Adjust to pH 2.5 (2.0–3.0 depending on use) with HCl.
- Bring to the final volume with water.
- Filter and store at 4 °C.
Good to know
- Collect each 1 mL fraction into 50–100 µL of 1 M Tris-HCl pH 8.5 to limit antibody denaturation.
FAQ
Can it strip western blots?
Yes — the mild stripping buffer (0.2 M glycine pH 2.2, 0.1% SDS, 1% Tween-20) works on the same principle.
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.