Usage and precautions of 4 - (2-hydroxyethyl) - 1-piperazine ethanesulfonic acid CAS NO.7365-45-9

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HEPES  (4-hydroxyethyl piperazine ethanesulfonic acid) is an zwitterionic buffer, CAS 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. It has no toxic effect on cells and can control the constant pH range for a long time. It can be used not only as a buffer in culture medium, but also as a pH regulator in cosmetics. HEPES is a non-ionic amphoteric buffer, which is at pH 7 2-7.4 good buffer capacity within the scope. Its major feature is to maintain a relatively stable pH value during open culture or cell observation. Under such culture standards, the cap of the cell culture bottle should be tightened to avoid a small amount of carbonate often needed in the culture medium from being dispersed into the air.

Usage of HEPES buffer:

① HEPES buffer can be immediately added to the configured culture medium according to the required concentration, and then filtered and sterilized. Add 2.38g HEPES to every 1000ml of culture medium, adjust the pH to 7.2 with lnnaoh after dissolution, and apply it after filtration and sterilization. At this time, the application concentration of HEPES is 10mmol / L.

② 100x storage solution (lmol / L) can also be prepared. Take 99ml culture solution and add LML storage solution before application. Finally, the concentration is still 10mmol / L. Configuration of lmol / L (100x) HEPES buffer: dissolve 23.8g HEPES in 90ml double distilled water, adjust the pH to 7.5-8.0 with lnnaoh, then use water titrant to 100ml, filter and sterilize, and store in bulk bottle (2ml / bottle) at 4 ℃ or - 20 ℃.

HEPES powder has high temperature resistance and melting point up to 200 ℃, so it will not degrade due to high-pressure sterilization. According to relevant literature, if HEPES aqueous solution is exposed to ambient light for three hours, it will produce cytotoxic hydrogen peroxide (H2O2). Therefore, it should be noted that:

1. HEPES aqueous solution must be kept away from light to avoid affecting the experimental results. After being prepared into solvent, it is recommended to store it at 4 ℃

The effect is better in the short term!

2. HEPES powder 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 buffer, with many kinds 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 you need anything, please contact us!