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HHBS (Hank's Buffer with Hepes) Preparation and Recipe

HHBS is a general purpose buffer that combines the stability of Hank's buffer with the good buffering capacity of HEPES buffering solution (one of Good's buffers). HHBS is used extensively for cell culture, but is sensitive to light.

Available for purchase here.

To prepare  L of HHBS (Hank's Buffer with Hepes):
Change the value in the textbox above to scale the recipe volume

Table 1. Required components
ComponentAmountConcentration
Calcium Chloride (mw: 110.98 g/mol)0.14 g0.00126 M
Magnesium Chloride Hexahydrate (mw: 203.30 g/mol)0.1 g0.00049 M
Magesium Sulfate Heptahydrate (mw: 246.47 g/mol)0.1 g0.00041 M
Potassium Chloride (mw: 74.55 g/mol)0.4 g0.00533 M
Potassium Phosphate Monobasic (mw: 136.09 g/mol)0.06 g0.00044 M
Sodium bicarbonate (mw: 84.01 g/mol)0.35 g0.00417 M
Sodium chloride (mw: 58.44 g/mol)8 g0.138 M
Sodium Phosphate Dibasic (mw: 141.96 g/mol)0.048 g0.00034 M
D-Glucose (Dextrose) (mw: 180.16 g/mol)1 g0.00556 M
Hepes (mw: 238.30 g/mol)4.76 g0.02 M
  1. Prepare 800 mL of distilled water in a suitable container.
  2. Add 0.14 g of Calcium Chloride to the solution.
  3. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution.
  4. Add 0.1 g of Magesium Sulfate Heptahydrate to the solution.
  5. Add 0.4 g of Potassium Chloride to the solution.
  6. Add 0.06 g of Potassium Phosphate Monobasic to the solution.
  7. Add 0.35 g of Sodium bicarbonate to the solution.
  8. Add 8 g of Sodium chloride to the solution.
  9. Add 0.048 g of Sodium Phosphate Dibasic to the solution.
  10. Add 1 g of D-Glucose (Dextrose) to the solution.
  11. Add 4.76 g of Hepes to the solution.
  12. Add distilled water until the volume is 1 L.

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Physiological Buffer
pH Buffering
Sample Preparation
BioAssays
Misc
Cell/Culture/Growth Media
Gel Electrophoresis


References

This online tool may be cited as follows

MLA

"Quest Calculate™ HHBS (Hank's Buffer with Hepes) Preparation and Recipe." AAT Bioquest, Inc.10 Aug2022https://www.aatbio.com/resources/buffer-preparations-and-recipes/hhbs-hanks-buffer-with-hepes.

APA

AAT Bioquest, Inc. (2022August 10). Quest Calculate™ HHBS (Hank's Buffer with Hepes) Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/hhbs-hanks-buffer-with-hepes.