HEPES buffer - an important buffer in cell culture

Details:

HEPES  (4-hydroxyethyl piperazine ethanesulfonic acid) is a hydrogen ion buffer with pH buffer range of 6.8-8.2. HEPES buffer is often indispensable in the process of cell culture. It is a weak acid. Under open culture conditions, if HEPES is added, it can prevent the oxidation of pH in the culture medium from rising, so as to maintain the pH at about 7.0. Let's continue to look at the application of HEPES in cell culture.

1. HEPES can maintain the structure and function of enzyme at low temperature

HEPES buffer is a weak acid, which is often used as zwitterionic buffer and pharmaceutical intermediate in organic chemical industry. The decomposition ability of the substance can be enhanced with the increase of temperature and weakened with the decrease of temperature. However, compared with other buffers, its decomposition constant changes little with temperature, which makes HEPES buffer a buffer that can better maintain the structure and function of the enzyme at low temperature.

2. HEPES pH value is beneficial to cell culture

Generally, the optimal culture environment for most cells is pH7 2-pH7. 4, and the buffer range of HEPES is just ph6 8-pH8. Within 2. This is beneficial to cell culture. Compared with other buffers, HEPES has higher stability in maintaining the pH value of cell culture medium, which 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.

3. HEPES is beneficial to the culture of Mycoplasma pneumoniae

HEPES ＾ is a commonly used buffer for cell culture. The final concentration is generally 10 ~ 50 ＾ mmol / L. generally, 20 ＾ mmol / L ＾ HEPES ＾ in the culture medium can serve as a buffer. The common concentration for cell culture is 10 ＾ mmol / L. HEPES buffer played a buffering role in this process, and the greater the concentration, the more obvious the buffering effect. When the medium pH ＾ was about ＾ 6.8 ＾ the cultured Mycoplasma pneumoniae entered the middle and late stage of logarithmic growth; The pH of the medium was 6.5 ~ 6.0, the content of Mycoplasma pneumoniae in the medium reached the plateau stage, and the bacterial quantity remained at a very high level.

4. HEPES is recommended for E. coli medium

The buffer system commonly used in E. coli culture medium is bicarbonate, which can provide nutrition, but it will reduce the buffer capacity under the condition of physiological pH value. HEPES is a strong buffer. Adding HEPES can make the cell culture medium at pH7 2-7.6 enhanced buffering capacity under conditions. The storage solution concentration of HEPES is generally 1m and stored at room temperature. The final concentration in the medium was 25mm, i.e. 5ml 1m HEPES was added to 200ml complete medium.

There are many important links in cell culture. A very important step is to select a suitable growth environment and high-quality culture medium for cells. For cell culture, it can not only ensure the stability and repeatability of your experimental data, but also save valuable experimental time and energy. HEPES produced by Desheng is white crystal powder, with main content ≥ 99.50%, iron ion ≤ 5ppm, sulfate ≤ 0.05%, and heavy metal residue (PB) ≤ 10ppm. Trial sample powder can be provided, which can be configured as a solution for testing.