AAT Bioquest

H-50 Buffer Preparation and Recipe

H-50 buffer is commonly used for washing and resuspending cells. It contains HEPES, one of the Good's zwitterionic buffers.

To prepare  L of H-50 Buffer:
Change the value in the textbox above to scale the recipe volume

Table 1. Required components
Hepes (mw: 238.30 g/mol)4.76 g0.02 M
Potassium Chloride (mw: 74.55 g/mol)3.73 g0.05 M
Magnesium sulfate (mw: 120.37 g/mol)0.12 g0.001 M
Sodium chloride (mw: 58.44 g/mol)0.58 g0.01 M
Sodium bicarbonate (mw: 84.01 g/mol)0.42 g0.005 M
Sodium phoshpate monobasic dihydrate (mw: 156.01 g/mol)0.156 g0.001 M
  1. Prepare 800 mL of distilled water in a suitable container.
  2. Add 4.76 g of Hepes to the solution.
  3. Add 3.73 g of Potassium Chloride to the solution.
  4. Add 0.12 g of Magnesium sulfate to the solution.
  5. Add 0.58 g of Sodium chloride to the solution.
  6. Add 0.42 g of Sodium bicarbonate to the solution.
  7. Add 0.156 g of Sodium phoshpate monobasic dihydrate to the solution.
  8. Add distilled water until the volume is 1 L.
  9. Adjust the pH to 7.0 with HCl or NaOH, as appropriate. Autoclave and store cold or frozen.

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


This online tool may be cited as follows


"Quest Calculate™ H-50 Buffer Preparation and Recipe." AAT Bioquest, Inc.16 Jun2024https://www.aatbio.com/resources/buffer-preparations-and-recipes/h-50-buffer.


AAT Bioquest, Inc. (2024June 16). Quest Calculate™ H-50 Buffer Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/h-50-buffer.