HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic organic chemical buffering agent. HEPES is widely used in cell culture, largely because it is better at maintaining physiological pH despite changes in carbon dioxide concentration (produced by cellular respiration) when compared to bicarbonate buffers, which are also commonly used in cell culture.

Application
HEPES has been used as a component of:
Hank’s balanced salt solution, Dulbecco’s modified eagle’s medium and no-glucose DMEM, which are used for the preparation of tissue slices.
Buffers used in localized conductance measurements on the basolateral side of culture inserts to examine cation selectivity.

General description:
HEPES has been described as one of the best all-purpose buffers available for biological research. At biological pH, the molecule is zwitterionic, and is effective as a buffer at pH 6.8 to 8.2 (pKa 7.55). It is typically used in cell culture at concentration between 5mM to 30 mM. HEPES has been used in a wide variety of applications, including tissue culture. It is commonly used to buffer cell culture media in air. HEPES finds its usage in in vitro experiments on Mg.

Hepes是一种氢离子缓冲剂，能较长时间控制恒定的pH范围，有效缓冲范围为pH6.8-8.2。可用于配制蛋白提取缓冲液、细胞培养用缓冲液等，对细胞无毒性作用。使用终浓度为10-50mmol/L，一般培养液内含20mmol/LHEPES即可达到缓冲能力，细胞培养常用终浓度为 5-30mM/L。