Using HEPES Buffer in Biochemistry and Molecular Biology Experiments

Biochemistry, Molecular Biology, and Cell Biology Protocols  >> Using HEPES Buffer in Biochemistry and Molecular Biology Experiments

HEPES buffer is a suitable alternative buffer when other buffers are not recommended for a certain application.  For example, Tris may react with NHS groups during biotinylation of NHS-biotin.  Calcium may precipitate with phosphates in phosphate-buffered saline buffers.  Therefore, HEPES is a suitable alternative in those cases.

Making 1M HEPES
Dissolve 238.3 g HEPES into 500 ml nanopure water
Stir, while adjusting the pH with NaOH or KOH to the appropriate pH (eg pH 7.4, pH 7.0)
Add nanopure water up to 1 litre
Filter if necessary
Store in the fridge if desired

Recipe for HBS (HEPES-buffered saline)
20 mM HEPES
150 mM NaCl

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