The biological buffer HEPES, 4-hydroxyethyl piperazine ethanesulfonic acid, is a hydrogen ion amphoteric buffer with good buffer capacity and long time to maintain pH constant. It is widely used in nucleic acid reaction buffer, hybridization buffer and cell culture medium. According to its characteristics, there are some differences in the configuration of buffer in different experiments.
Physical and chemical properties of HEPES
HEPES mother liquor (buffer stock): directly prepare 0.5-1m HEPES solution and NaOH solution, control the mixing ratio of the two to adjust the pH, and then use distilled water or ultra pure water to fix the volume. If necessary, NaOH can be replaced by KOH.
HEPES buffer salt solution: add a small amount of sodium chloride and disodium hydrogen phosphate into HEPES solution, then add NaOH aqueous solution, adjust pH, add distilled water for constant volume, and store at low temperature.
HEPES cell culture medium: add a small amount of sodium chloride, potassium chloride, disodium hydrogen phosphate, dextran, etc. into HEPES aqueous solution, then adjust the pH with NaOH solution, and finally store at a constant volume and low temperature. In cell adhesion experiments, calcium and magnesium ions are usually added to HEPES culture medium to protect the cadherin of cells and not affect the formation of cell aggregation.
HA solution: hepes-bsa cell culture medium, which is one of the components of cell culture medium, does not contain calcium and magnesium ions, and contains some other salt ions. After the treatment of filtration bacteria, it is mostly suitable for cleaning and culture in primary culture of tissue cells.
The above is the application difference of HEPES buffer developed and produced by Desheng in different experiments. In addition, the non inactivated virus preservation solution newly developed by the company also contains HEPES buffer. Because it has no toxic effect on cells, it not only maintains pH, but also increases the survival time of virus host cells after sampling in vitro, retains the integrity of virus nucleic acid and protein to the greatest extent, and improves the detection accuracy.