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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.

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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)##
Magnesium Chloride Hexahydrate (mw: 203.3 g / mol)##
Magesium Sulfate Heptahydrate (mw: 246.47 g / mol)##
Potassium Chloride (mw: 74.55 g / mol)##
Potassium Phosphate Monobasic (mw: 136.09 g / mol)##
Sodium bicarbonate (mw: 84.01 g / mol)##
Sodium chloride (mw: 58.44 g / mol)##
Sodium Phosphate Dibasic (mw: 141.96 g / mol)##
D-Glucose (Dextrose) (mw: 180.16 g / mol)##
Hepes (mw: 238.3 g / mol)##
  • Prepare 800 mL of distilled water in a suitable container.
  • Add # of Calcium Chloride to the solution
  • Add # of Magnesium Chloride Hexahydrate to the solution
  • Add # of Magesium Sulfate Heptahydrate to the solution
  • Add # of Potassium Chloride to the solution
  • Add # of Potassium Phosphate Monobasic to the solution
  • Add # of Sodium bicarbonate to the solution
  • Add # of Sodium chloride to the solution
  • Add # of Sodium Phosphate Dibasic to the solution
  • Add # of D-Glucose (Dextrose) to the solution
  • Add # of Hepes to the solution
  • Add distilled water until the volume is 1 L.


References
This online resource may be cited as follows
MLA
"Quest CalculateHHBS (Hank's Buffer with Hepes) Preparation and Recipe." AAT Bioquest, Inc.9 Oct2025https://www.aatbio.com/resources/buffer-preparations-and-recipes/hhbs-hanks-buffer-with-hepes.
APA
AAT Bioquest, Inc. (2025October 9). Quest CalculateHHBS (Hank's Buffer with Hepes) Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/hhbs-hanks-buffer-with-hepes.
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This online resource has been cited in 1 publications, including
Identification of Autophagy as a Functional Target Suitable for the Pharmacological Treatment of Mitochondrial Membrane Protein-Associated Neurodegeneration (MPAN) In Vitro
Authors: 
Zanuttigh, Enrica and Derderian, Kevork and G{\"u}ra, Miriam A and Geerlof, Arie and Di Meo, Ivano and Cavestro, Chiara and Hempfling, Stefan and Ortiz-Collazos, Stephanie and Mauthe, Mario and Kmie{\'c}, Tomasz and others,
Journal: 
Pharmaceutics (2023): 267