The use and preparation method of HEPES buffer

Details:

Hepes is a non-ionic two-component buffer, the main component of which is hydroxyethylpiperazine ethanesulfonic acid. It has a good buffer capacity in the range of PH7.2-7.4. Hepes has a wide range of applications. When used as a biological buffer, it can be used as a buffer reagent in cell culture media, as a buffer and eluent in protein research, and can also be used in DNA/RNA extraction kits and PCR diagnostic kits.

When Hepes is used as a buffer, it can maintain a relatively constant pH value during open culture or cell observation. Under such culture conditions, the cell culture device needs to be closed and perfect to avoid the small amount required in the culture medium. The carbonate disperses into the air. There are also two ways to use Hepes buffer. details as follows:

1. Hepes can be directly added to the prepared culture solution according to the required concentration, and then filtered and sterilized. Add 2.38 grams of HEPES to each 1000ml culture solution, and adjust the pH to 7.2 with 1N NaOH after dissolving, and use after filtering and sterilization At this time, the use concentration of HEPES is 10 mmol/L.
2. It can also be made into 100 x stock solution (l mol/L). Before use, take 99ml culture solution and add 1ml stock solution, the final application concentration is still 10mmol/L. l mol/L (100 x) HEPES stock solution preparation method: Dissolve 23.8g HEPES in 90ml double-distilled water, adjust the pH to 7.5-8.0 with 1N NaOH, then dilute to 100ml with water, filter and sterilize, and divide into vials ( 2ml/bottle), need to be stored at 4℃ or -20℃.

In daily biochemical experiments, especially during cell culture, Hepes buffer is usually indispensable. And the use of Hepes buffer has been understood, and we will learn about the preparation method of Hepes together later.
Hepes preparation method: directly use 1,2-dichloroethane as the solvent, add hydroxyethylpiperazine (5.00g, 0.02mol), potassium carbonate K2CO3 (6.00 g, 0.04mol), 50mL 1,2-dichloroethane, heated in an oil bath at 90°C (1,2-dichloroethane boiling point 85°C), and stirred for reaction for 20h. The reaction was stopped, filtered, and the filtered salt was washed with 200 mL of ethyl acetate (EA). The filtrate was spin-dried to obtain 2.6 g of HEPES solid.

In summary, the application of Hepes buffer is very wide, and the method of use and preparation are not complicated. This has led to many Hepes merchants in the market, and there are actually only a few real domestic manufacturers. Therefore, the market is currently mixed, with many middlemen earning the difference. Recommend a manufacturer here, Hubei Xindesheng Materials Co., Ltd., Desheng Company has been specialized in the production and research of biological buffers for many years. It has multi-level factories and imported equipment, and has created a professional team responsible for R&D and production management, with high production efficiency , The product quality is excellent, the warehouse stock is sufficient, and the order is shipped immediately. If you are interested in knowing about Desheng’s products, you can contact customer service on Desheng’s official website for details.