Characteristics, application and storage recommendations of HEPES buffer

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HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid) is a zwitterionic buffer with CAS number 7365-45-9. This buffer is usually used in biochemical experiments. To adjust the pH of the reaction system, this component is contained in virus preservation solutions or cell preservation solutions. It is non-toxic to cells and can control a constant pH range for a long time. It can not only be used as a buffer in the culture medium, but also as a pH regulator in cosmetics.


·The packing specification of HEPES buffer is 25kg/barrel

The characteristics of a good HEPES buffer:

1. The pKa value is between 6.0 and 8.0;

2. Has high solubility in water;

3. It has membrane impermeability and is not easy to penetrate biological membranes;

4. Limited impact on biochemical reactions, resistance to chemical and enzymatic hydrolysis, and does not form complexes or precipitates with metal ions;

5. Has extremely low visible light and ultraviolet light absorption;

6. Ion concentration, solution composition and temperature have little effect on dissociation;

HEPES handling and storage recommendations

HEPES powder is resistant to high temperatures and has a melting point of up to 200°C, so it will not be degraded by autoclaving. According to related literature, if the HEPES aqueous solution is exposed to ambient light for three hours, cytotoxic hydrogen peroxide (H2O2) will be produced. Therefore, it should be noted that the HEPES aqueous solution must be kept away from light to avoid affecting the experimental results. The HEPES powder is generally placed in a dry room when placed, and cannot be exposed to direct sunlight for a long time

Application of HEPES in cell culture

HEPES is widely used in a variety of biochemical reactions and is used as a buffer reagent in some cell culture media. The final concentration of HEPES buffer is 10-50mmol/L. When applied to cell culture, the general culture medium contains 20mmol/L LHEPES can reach the buffering capacity. HEPES does not directly supply nutrients to the cells. In open culture conditions, adding HEPES buffer can prevent the pH from oxidizing in the medium to increase, thereby maintaining the pH at about 7.0.

Desheng Technology is a professional enterprise engaged in the research, production and sales of scientific research and biochemical reagents. The company adopts advanced production technology and equipment, and a humanized office environment. It has gathered a group of energetic and pragmatic technical personnel and a strict matrix management team. The HEPES currently provided by Desheng is a powder reagent, and it can be used if required. Formulated into a ready-to-use buffer solution of a specific concentration.