Hank’s Balanced Salt Buffer Protocol (HBSS Buffer)

Summary of use

Hank’s Balanced Salt Solution (HBSS) is a buffer solution widely used in cell biology research experiments, cell washing so, cell separation, cell/tissue transferring with combination of phenol red. HBSS Buffer is rich in bicarbonate ions and well known as its non-Reactive properties. Hank’s Balanced Salt Solution (HBSS) has been used in immunohistochemistry rinsing when targe molecule is not phosphatase. Hank’s Balanced Salt Solution (HBSS) is a sensitive buffer to storage. It is better to make and use freshly.

Hank’s Balanced Salt Buffer Protocol (HBSS Buffer)

Preparation of Hank’s Balanced Salt Buffer Solution (HBSS Buffer)

1. 1. 1. Prepare 950 mL of distilled water in a suitable container.
      2. Add 8 g of Sodium chloride to the solution.
      3. Add 0.4 g of Potassium Chloride to the solution.
      4. Add 0.14 g of Calcium Chloride to the solution.
      5. Add 0.1 g of Magesium Sulfate Heptahydrate to the solution.
      6. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution.
      7. Add 0.06 g of Sodium Phosphate Dibasic Dihydrate to the solution.
      8. Add 0.06 g of Potassium Phosphate Monobasic to the solution.
      9. Add 1 g of D-Glucose (Dextrose) to the solution.
      10. Add 0.35 g of Sodium bicarbonate to the solution.
      11. Add distilled water until the volume is 1 L.

Note1: For sterilization you need to filter Hank’s Balanced Salt Solution (HBSS) with 0.22 μm sterile filter.

Note 2: Keep Hank’s Balanced Salt Solution refrigerated. It is stable for 3-5 days.