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|Chemical Name:||4-(2-Hydroxyethyl)-1-Piperazineethanesulfonic Acid||MF:||C8H18N2O4S|
CAS No 7365-45-9,
7365-45-9 HEPES Buffer Solution,
White Crystalline HEPES Buffer Solution
Preparation and application of 2 - (4 - (2-hydroxyethyl) piperazin-1-yl) ethanesulfonic acid buffer
In the process of cell culture, HEPES is often indispensable. In open culture conditions, if HEPES is added, it can prevent the increase of pH oxidation in the culture medium, so as to maintain the pH at about 7.0, effectively improve the buffer capacity of virus culture medium, provide a suitable environment for the virus, and promote the stability of virus particles. In addition, it can better maintain the structure and function of the enzyme at low temperature . How about the preparation and use of HEPES buffer? Let's give you a brief introduction.
|Melting point||234-238 ° C|
|Density||1.07g/mlat 20 ° C|
|Storage condition||2-8 ° C|
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 500 ml 1 m HEPES, pH = 7.0, stock solution
Dissolve 119.15 g 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 at 4 ℃.
2, HEPES buffer formula with small amount of salt (500ml)
HEPES 6.5g, NaCl 8.0g, na2hpo4.7h2o 0.198g, adjust pH value with 0.5m NaOH solution, * and then fix volume.
3, Preparation of 2 × HEPES buffer salt solution
Dissolve 1.6g NaCl, 0.074g KCl, 0.027g na2hpo4.2h2o, 0.2g 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.
There are two ways to use HEPES
(1) HEPES can be directly added into the prepared culture medium according to the required concentration, and then filtered to remove bacteria. Add 2.38g HEPES into every 1000ml culture medium, adjust pH to 7.2 with LN NaOH after dissolving, filter and use after sterilization. The concentration of HEPES was 10 mmol / L.
(2) It can also be made into 100 x storage solution (L mol / L). Before use, add 99 ml culture solution into 1 ml storage solution, * the final application concentration is still 10 mmol / L. Methods: 23.8g HEPES was dissolved in 90ml double distilled water, adjusted to pH 7.5-8.0 with LN NaOH, and then diluted to 100ml with water, filtered and sterilized, sub packed into small bottles (2ml / bottle), stored at 4 ℃ or - 20 ℃.
The final concentration of HES is 50 mmol / L, and it needs to be noted that PES is non-toxic to cells. In general, the buffer capacity of HEPES with a concentration of 20mmol / l can be achieved, and it is OK for us to prepare it correctly and flexibly according to the actual situation.
Now the specification of HEPES sold by Desheng is 25kg / barrel, which can also be packed according to customers' purchase quantity. The larger the order quantity, the more favorable the price. Welcome to inquire and order!
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