Preparation steps of HEPES buffer

Details:

HEPES  chemical name: 4-hydroxyethyl piperazine ethanesulfonic acid, cas7365-45-9. HEPES is commonly used as biological buffer, and can also be used to stabilize pH value in cosmetics, coatings, surfactants, etc. The pH buffer range is 6.8-8.2, which is used in biochemical diagnostic kit, DNA / RNA extraction kit and PCR diagnostic kit. It has no toxic effect on cells. It is a hydrogen ion buffer, which can control the constant pH range for a long time.

HEPES buffer is often used in cell culture experiments. In the open culture condition, adding HEPES buffer can prevent the increase of pH oxidation in the culture medium, so as to maintain the pH at about 7.0. Then, what are the preparation steps and methods of HEPES buffer Next, Desheng will introduce it in detail.

Different literatures have different opinions on the preparation of HEPES buffer solution. According to the use, one is pure HEPES + NaOH and the other is HEPES + salt. Various preparation methods are summarized as follows:

1、 Preparation of 500ml 1 m HEPES, pH = 7.0 stock solution

Dissolve 119.15 g of HEPES in 400ml distilled water, add 0.5 ~ 1m NaOH aqueous solution to adjust at least the required pH (the effective pH range of HEPES is 6.8 ~ 8.2), then fix the volume to 500ml with distilled water and store it at 4 ℃.

2、 HEPES buffer formula with a small amount of salt (500ml)

HEPES 6.5g, NaCl 8.0g, na2hpo4.7h2o 0.198g, adjust the pH value with 0.5m NaOH aqueous solution, and finally fix the volume.

3、 2 × Preparation of HEPES buffer salt solution

Dissolve 1.6g NaCl, 0.074g KCl, 0.027g na2hpo4.2h2o, 0.2g glucan or dextran and 1g HEPES in 90ml distilled water, adjust to the required pH value with 0.5m NaOH, and then fix the volume to 100ml with distilled water.

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