Western blot stripping buffer (mild) recipe
A mildly acidic buffer that removes antibodies so a membrane can be re-probed.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Glycine | 7.5 g | 200 mM |
| SDS | 500 mg | 0.1% |
| Tween-20 | 5 mL | 1% |
| Water (distilled) | to final volume |
Steps
- Dissolve glycine and SDS in about 800 mL of water and add Tween-20.
- Adjust to pH 2.2 with HCl and make up to 1 L.
- Incubate the membrane twice for 5–10 min at room temperature, wash well with PBS/TBST and block again.
Good to know
- Some protein is lost at every strip — avoid quantitative comparisons before vs after stripping.
- Check stripping by re-applying ECL alone and confirming there is no signal.
FAQ
What if antibodies do not come off?
Harsh stripping with β-ME and SDS at 50 °C works but loses more protein; PVDF tolerates repeated stripping better.
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.