Application of buffer HEPES in cell culture medium CAS NO.7365-45-9

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When should HEPES be used in cell culture medium

HEPES (4-hydroxyethyl piperazine ethanesulfonic acid) is a commonly used zwitterionic buffer with a pKa of 7.3 at 37 ° C. In addition to sodium bicarbonate, we should also use HEPES in the culture medium, because it is important to maintain sufficient bicarbonate in the culture medium to achieve nutritional purposes. If necessary, HEPES can be added to the cell culture medium at a final concentration of 10 mm to 25 mm to provide a balance External buffer capacity. Lower concentrations are usually insufficient to control pH fluctuations, while higher concentrations are usually toxic.

Since the buffer capacity of HEPES is independent of CO2 concentration, it is an ideal buffer incubator to maintain the culture pH when cell culture needs to be operated outside CO2 for a long time. HEPES buffer medium can resist sharp pH changes, but can not completely prevent pH changes. Since high levels of HEPES can cause cytotoxicity, if there is significant toxicity to specific cell lines or primary cell cultures, the concentration of this organic buffer should be reduced.

Why use HEPES

Because the growth and metabolism of cells in culture will lead to drastic changes in the environmental pH value of the culture medium, it is necessary to add an appropriate amount of biological buffer to maintain the stable environment of cultured cells. However, there are so many biological buffers and substitutes on the market for cell culture. Why use HEPES

Generally, the Zui best culture environment for most cells is pH7 2-pH7. 4. The buffer range of HEPES is just ph6 8-ph 8.2, which is favorable for cell culture. Compared with other buffers (such as PBS (phosphate buffered saline) and Tris), HEPES has higher stability in maintaining the pH value of cell culture medium. This is also the reason why HEPES is widely used in cell culture, tissue culture, protein purification and extraction, immunoprecipitation, cell lysis, live cell imaging and other biological / biochemical studies.

What are the benefits of using HEPES

1. HEPES acts as a buffer for enzyme at low temperature

Generally, the use condition of biological buffer is 25 ℃ at room temperature, Or higher temperatures (according to the optimum pH range of the enzyme), but sometimes experiments need to be carried out under low temperature conditions, and some buffers are obviously affected by low temperature and are not suitable for use. The decomposition ability of HEPES can increase with the increase of temperature and decrease with the decrease of temperature. However, compared with other buffers, its decomposition constant changes little with temperature, which makes HEPES buffer can better maintain the knot of the enzyme at low temperature Buffer of structure and function.

2. HEPES has no toxic and side effects on cells

HEPES buffer can control the constant pH range for a long time. Low concentration HEPES has no toxic effect on cells. It is often used in cell culture. For example, HEPES can be added to the culture solution or virus preservation solution of various bacteria and fungi to maintain the pH environment required by cells, bacteria and viruses.