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

Component	Amount	Concentration
Hepes (mw: 238.3 g / mol)	#	#
Potassium Chloride (mw: 74.55 g / mol)	#	#
Magnesium sulfate (mw: 120.37 g / mol)	#	#
Sodium chloride (mw: 58.44 g / mol)	#	#
Sodium bicarbonate (mw: 84.01 g / mol)	#	#
Sodium phoshpate monobasic dihydrate (mw: 156.01 g / mol)	#	#

Prepare 800 mL of distilled water in a suitable container.
Add # of Hepes to the solution
Add # of Potassium Chloride to the solution
Add # of Magnesium sulfate to the solution
Add # of Sodium chloride to the solution
Add # of Sodium bicarbonate to the solution
Add # of Sodium phoshpate monobasic dihydrate to the solution
Add distilled water until the volume is 1 L.
Adjust the pH to 7.0 with HCl or NaOH, as appropriate. Autoclave and store cold or frozen.

References

This online resource may be cited as follows

MLA	"Quest Calculate™ H-50 Buffer Preparation and Recipe." AAT Bioquest, Inc., 5 Nov. 2025, https://www.aatbio.com/resources/buffer-preparations-and-recipes/h-50-buffer.
APA	AAT Bioquest, Inc. (2025, November 5). Quest Calculate™ H-50 Buffer Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/h-50-buffer.
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