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Buffering Features

HEPES is a zwitterionic sulfonic acid buffering agent. It was one of the first Good’s buffers described by Norman E. Good in 1966.

HEPES has the following characteristics:

• pKa1 (25 °C) = 3

• pKa2 (25 °C) = 7.5

• Useful pH range = 2.5 to 3.5 or 6.8 to 8.2

Applications

Compared with other buffers such as PBS (phosphate buffered saline) and TRIS, HEPES has higher stability in maintaining the pH values of the cell culture media, that's also the reason why HEPES is widely used in cell culture , tissue culture, protein purification and extraction, immunoprecipitation, cell lysis, live.

HEPES has negligible metal ion binding, so it is a good choice of buffers for enzymes that may be inhibited by metal chelation.

Tips

HEPES is very soluble in water

Affects the membrane potential of neurons

HEPES rarely binds to metal ions

Keep HEPES-containing solutions in darkness as much as possible to prevent oxidation