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Wuhan Desheng Biochemical Technology Co., Ltd
Wuhan Desheng Biochemical Technology Co., Ltd
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The Application of HEPES Buffer

The Application of HEPES Buffer

The Application of HEPES Buffer


    HEPES (4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic buffer, CAS No. 7365-45-9, this buffer is generally used in biochemical experiments to adjust the PH of the reaction system, such as virus preservation solution or cell preservation solution. It has no toxic on cells,and maintain PH for a long time, It is always applied in cell culture medium and cosmetics.

HEPES is a non-ionic amphoteric buffer with good buffering capacity in the pH range of 7.2-7.4. Its major feature is to maintain a relatively stable PH value in culture medium or cell observation. Under these standards, the caps of cell culture flasks should be tightened tightly to avoid required carbonate escaping into the air.


The way to prepare the HEPES buffer.

    Hepes buffer can be added directly into the prepared culture medium with required concentration, followed by filter sterilization.Add 2.38 grams of HEPES powders to every 1000ml of culture solution, adjust the PH to 7.2 with 1NNaOH after dissolving, filter and sterilize before applying, at this time, the concentration of HEPES is 10mmol/L.

    Adding 99ml cell culture medium solvent into 1ml stock solution,the concentration of HEPES is also 10mol/L.Below is the way to prepare HEPES stock solution with 1mol/L. First, dissolving 23.8g HEPES into 90ml double-distilled water;Second,adjusting the pH to 7.5-8.0 with 1N NaOH; Third,making up to 100ml with water titrant;Forth,filtering and sterilizing before packed into 2ml bottle;At last stored at 4℃ or -20℃.

With high temperature resistant and melting point up to 200°C,HEPES powder won’t be degraded by autoclave. According to relevant literature, if the HEPES aqueous solution is exposed to ambient light for three hours, toxic hydrogen peroxide (H2O2) will be generated. Therefore,2 points need to be paid attention to,one is the HEPES aqueous solution must be kept away from light so as not to affect the experimental results. After being configured as a solvent, it should be stored at 4 °C and used in a short period of time for better results.Another is generally placed in a dry room and cannot be directly exposed to sunlight for a long time.



    Hubei New Desheng Material Technology Co.,Ltd is specialized in producing biological Buffers over years.We’ve developed a series of biological buffers including TRIS, HEPES, TAPS, MOPS, CAPS, BICINE, EPPS, PIPES and so on. It’s not only have a large market share in the domestic market, but also have been sold to dozens of countries around the world. We’ve established long-term and stable cooperation with many large-scale biomedical technology enterprises.

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