HEPES (hydroxyethyl-piperazineethane-sulfonic acid buffer) is an organic buffer and very stable. In the physiological area HEPES has an excellent buffer capacity in a pH range of 7.2 to 7.6. HEPES is apt for all cell culture systems with a high proliferation and cell density. On HEPES buffered cell culture media, the incubation in a CO2 incubator is not necessary.
How to use HEPES buffer:
- • dissolve powder and adjust pH at 7.2 with sodium hydroxide
- • add 10-20 ml of HEPES buffer per litre of medium (for very delicate cell cultures, use only 10 ml)
- • a concentration of 20 mM HEPES requires 20 ml/l medium, for 25 mM use 25 ml/l
- • if NaHCO3 is required, only add 10 %
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