The use and precautions of Hepes buffer CAS NO.1185-53-1

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HEPES (4-Hydroxyethylpiperazineethanesulfonic acid) is a zwitterionic buffer, CAS No. 7365-45-9, this buffer is usually used in biochemical experiments to adjust the pH value of the reaction system, Such as virus preservation solution or cell preservation solution contains this component. It has no toxic effect on cells, can control a constant pH range for a long time, and can not only be used as a buffer in the culture medium, but also as a pH regulator in 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 during open culture or cell observation. Under such culture standards, the caps of cell culture flasks should be tightened tightly to avoid escaping into the air the small amount of carbonate that is often required in the culture medium.
How to use hepes buffer:
①Hepes buffer can be added to the prepared culture solution immediately according to the required concentration, and then filtered and sterilized. Add 2.38 grams of HEPES to every 1000 ml of culture solution, adjust the pH to 7.2 with 1N NaOH after dissolving, filter and sterilize before applying, at this time, the application concentration of HEPES is 10 mmol/L.
②You can also prepare 100x stock solution (1mol/L), take 99ml of culture medium and add 1ml of stock solution before application, and the final application concentration is still 10mmol/L. lmol/L (100x) hepes buffer configuration: dissolve 23.8g of HEPES in 90ml of double-distilled water, adjust the pH to 7.5-8.0 with 1N NaOH, then make up to 100ml with water titrant, filter and sterilize, bulk bottles (2ml/bottle), 4 Store at ℃ or -20℃.
HEPES powder is resistant to high temperatures and has a melting point of up to 200°C, so it will not degrade by autoclaving. According to the relevant literature, if an aqueous solution of HEPES is exposed to ambient light for three hours, cytotoxic hydrogen peroxide (H2O2) will be produced. Therefore, care should be taken:
1. 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. It is recommended to use it in a short period of time for better results!
2. When HEPES powder is placed, it is generally placed in a dry room and cannot be directly exposed to sunlight for a long time.
Desheng Biochemical is a professional manufacturer of biological buffers, with many types of products. Since its establishment in 2005, it has more than ten years of R&D and production experience. The HEPES buffer produced by Desheng has high purity and wide application range. If necessary, please contact us!