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Preparation method of HEPES buffer

Update Time:2020/3/16 13:30:52   View times:

HEPES solution: a weak acid, English name is 4- (2-hydroxyerhyl)piperazine-1-erhanesulfonic acid. It can be used as biological buffer reagent to prepare protein extraction buffer and cell culture buffer. PH range: 6.8-8.2.

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

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

119.15 g HEPES are dissolved in 400ml distilled water, and 0.5-1m NaOH solution is added to adjust at least the required pH (the effective pH range of HEPES is 6.8-8.2), then the volume is fixed to 500ml with distilled water and stored at 4 ℃.

2、 HEPES buffer 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、 Preparation of 2 × 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 of distilled water to 100ml.


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