HBSS (Hanks' balanced salt solution, Ca²⁺/Mg²⁺-free) recipe
A glucose-containing salt solution for washing and handling cells and tissue in room air (outside the CO₂ incubator).
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| NaCl | 4 g | 137 mM |
| KCl | 200 mg | 5.4 mM |
| KH₂PO₄ | 30 mg | 0.44 mM |
| Na₂HPO₄ (anhydrous) | 24 mg | 0.34 mM |
| D-glucose | 500 mg | 5.6 mM |
| NaHCO₃ | 175 mg | 4.2 mM |
| Water (distilled) | to final volume |
Steps
- Dissolve in about 80% of the final volume of water.
- Check pH 7.2–7.4.
- Bring to the final volume with water.
- Filter-sterilise through 0.22 µm. Store at 4 °C.
Good to know
- HBSS with Ca²⁺/Mg²⁺: add CaCl₂ 0.14 g, MgCl₂·6H₂O 0.1 g and MgSO₄·7H₂O 0.1 g per litre (dissolve separately, add last).
⚠ Safety Handle solutions for live cells aseptically in a biosafety cabinet.
FAQ
How is it different from PBS?
It contains glucose and a little bicarbonate, so cells tolerate it longer — but it is not a culture medium.
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.