Western blot transfer buffer (Towbin) recipe
For wet transfer of proteins from gel to membrane: 25 mM Tris, 192 mM glycine, 20% methanol.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Tris base | 3.03 g | 25 mM |
| Glycine | 14.4 g | 192 mM |
| Methanol | 200 mL | 20% (v/v) |
| Water (distilled) | to final volume |
Steps
- Dissolve Tris and glycine in about 700 mL of water.
- Add methanol and make up to 1 L.
- Chill to 4 °C before use.
Good to know
- For large proteins (>150 kDa), reduce methanol to ≤10% and add 0.0375–0.1% SDS.
- For a 10X stock, concentrate only Tris and glycine and add methanol when diluting.
⚠ Safety Methanol is toxic and flammable — work with ventilation.
FAQ
Why add methanol?
It strips SDS so proteins bind the membrane and stops the gel swelling — but it slows the elution of large proteins.
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.