Carbonate-bicarbonate coating buffer (pH 9.6, ELISA) recipe
Coats capture antibody or antigen onto ELISA plates: 0.05 M carbonate-bicarbonate, pH 9.6.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| Na₂CO₃ (anhydrous) | 159 mg | 15 mM |
| NaHCO₃ | 293 mg | 35 mM |
| Water (distilled) | to final volume |
Steps
- Dissolve in about 80% of the final volume of water.
- Check pH 9.6 (usually no adjustment needed).
- Bring to the final volume with water.
- Store at 4 °C and use within 1–2 weeks.
Good to know
- Dilute antibody to 1–10 µg/mL, add 50–100 µL per well and coat overnight at 4 °C.
- Some antibodies coat better in PBS — check the datasheet.
FAQ
What if it sits open for long?
It absorbs CO₂ and the pH drops — keep it capped.
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.