Use and preparation of HEPES buffer

Details:

 HEPES  is a non-ionic buffer, and its main component is hydroxyethyl piperazine ethanethiosulfonic acid. It is at pH7 2-7.4 it has good buffer capacity in the range of. HEPES is widely used. As a biological buffer, it can be used in cell culture medium as buffer reagent, as buffer and eluent in protein research, as well as DNA / RNA extraction kit and PCR diagnostic kit.

HEPES as a buffer has a very good point, that is, it can maintain a relatively constant pH value during open culture or cell observation. Under this culture condition, the cell incubator needs to be closed and perfect to avoid a small amount of carbonate required in the culture medium from being dispersed into the air. There are two ways to use HEPES buffer. The details are as follows:

1. HEPES can be directly added to the prepared 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 LN NaOH after dissolution, and use after filtration and sterilization At this time, the concentration of HEPES is 10 mmol / L

2. It can also be prepared into 100 x storage solution (L mol / L). Before use, 99ml culture solution is added into LML storage solution, and the final application concentration is still 10mmol / L. preparation method of L mol / L (100 x) HEPES storage solution: dissolve 23.8g HEPES in 90ml double distilled water, adjust the pH to 7.5-8.0 with LN NaOH, then fix the volume with water to 100ml, filter and sterilize, sub pack small bottles (2ml / bottle), and store them at 4 ℃ or - 20 ℃

HEPES buffer is usually indispensable in daily biochemical experiments, especially in cell culture. The use method of HEPES buffer has been understood. Later, let's learn about the preparation method of HEPES.

Preparation method of HEPES: directly use 1,2-dichloroethane as solvent and add hydroxyethyl piperazine (5.00g, 0.02mol) into a 100ml three mouth bottle equipped with mechanical stirring and thermometer, Potassium carbonate K2CO3 (6.00g, 0.04mol), 50ml1,2-dichloroethane, heated in oil bath at 90 ℃ (boiling point of 1,2-dichloroethane at 85 ℃), stirred for reaction for 20h. Stop the reaction, filter, and wash the filtered salt with 200ml ethyl acetate (EA). Spin the filtrate dry to obtain 2.6g HEPES solid.

In conclusion, HEPES buffer is widely used, and the application method and preparation method are not complex. This leads to many HEPES businesses in the market, and there are only a few real domestic manufacturers at present. Therefore, the market is mixed at present, and many middlemen earn the price difference. We recommend a manufacturer here, Hubei xindesheng Material Co., Ltd. Desheng company has been specializing in the production and R & D of biological buffer for many years, has multi-storey plants and imported equipment, and has established a professional team to be responsible for R & D and production management. It has high production efficiency, excellent product quality and sufficient stock in the warehouse. It can be delivered immediately after placing an order. If you want to know about Desheng's products, you can contact customer service on Desheng's official website for details.