Application of biological buffer HEPES

Details:

HEPES  chemical name: 2 - [4 - (2-hydroxyethyl) - 1-piperazinyl] ethanesulfonicacid Chinese Name: hydroxyethyl piperazine ethylthiosulfonic acid molecular formula: c8h18n2o4shepes. It is an amphoteric organic chemical buffer with no toxic effect on cells. At the same time, it is also a hydrogen ion buffer, which can control the constant pH range for a long time, It has good buffering capacity in the pH value range of 7.2-7.4.

A good HEPES buffer has the following characteristics:

1. PKa values are between 6.0 and 8.0

2. High solubility

3. Membrane impermeability

4. Limited impact on biochemical reaction

5. Very low visible and ultraviolet absorption

6. Resistance to chemical and enzymatic hydrolysis

7. Easy to prepare

HEPES is widely used in a variety of biochemical reactions and as a buffer reagent in some cell culture media. Its biggest advantage is that it can maintain a constant pH value in open culture or cell observation. The concentration of HEPES in the cell culture medium ranges from 10 mm to 25 mm. Note that HEPES does not provide nutrition for cells. Only when cells need to be treated outside the CO2 incubator for a long time in the process of cell culture can they be added to the culture medium to enhance the buffer capacity.

HEPES can be used in two ways:

1. Add directly into the configured culture solution according to the required concentration, filter and sterilize. Add 2.38g HEPES per 1000ml of culture solution. After dissolution, adjust the pH to 7.2 with 1n NaOH. It can be used after filtration and sterilization. At this time, the concentration of HEPES is 10mmol / L.

2. Prepare 100x storage solution (1mol / L), dissolve 23.8g HEPES in 90ml double distilled water, adjust the pH to 7.5-8.0 with 1nnaoh, then fix the volume with water to 100ml, filter and sterilize, divide into small bottles, and store at 4 ℃ to 20 ℃. Before use, take 99ml culture solution and add 1ml storage solution, and the final application concentration is still 10mmol / L

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