5% non-fat milk blocking solution recipe
Covers empty sites on western blot membranes to lower background: 5% non-fat dry milk in TBST.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Non-fat dry milk | 2.5 g | 5% (w/v) |
| TBST | as needed | to final volume |
Steps
- Add milk powder to TBST and shake or stir until dissolved (10–20 min).
- Make fresh; leftovers keep 1–2 days at 4 °C.
Good to know
- Use 5% BSA for phospho-specific antibodies or biotin detection (milk contains casein and biotin).
- Undissolved particles can leave spots — filter if needed.
FAQ
Can I dilute antibodies in blocking solution?
Usually yes — but dilute phospho-specific antibodies in BSA.
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.