HEPES is a kind of nonionic amphoteric buffer. The main component of HEPES buffer is 2 - [4 - (2-hydroxyethyl) - 1-piperazinyl] ethanesulfonic acid. HEPES is a kind of nonionic amphoteric buffer with good buffering capacity in the range of pH 7.2-7.4. In this condition, the lid of the cell culture bottle should be tightened to prevent a small amount of carbonate needed in the culture medium from scattering into the air.
Most of the culture media contain phenol red as pH indicator. The acidic culture media are orange yellow, and the alkaline culture media are dark red. HEPES is expensive and may be toxic to some cells at high concentrations, HEPES can be used as buffer instead of bicarbonate to remove the limitation of high concentration of CO2 culture environment. In practice, it is not so simple. Dissolved CO2 and bicarbonate are also very important for good cell growth.
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) Before use, 99 ml of culture medium was added to 1 ml of storage medium, and the final concentration was 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 ℃.
4-hydroxyethyl piperazine ethanesulfonic acid has no toxic effect on cells. It is a hydrogen ion buffer, which can control the constant pH range for a long time. When the final concentration is 10-50 mmol / L and the culture medium contains 20 mmol / L HEPES, the buffer capacity can be achieved. It can be used as biological buffer, reaction buffer, pre hybridization buffer and hybridization buffer for separation and analysis of RNA nuclear components, and for RNA and t4rna.
Molecular biology grade is used for RNA3 '- end labeling with t4rna ligase chemicalbook, separation and analysis of reaction buffer components, pre hybridization buffer and buffer components of nuclear RNA. Cell to cell adhesion, short-term cell aggregation culture, tissue and cell cleaning buffer.
Desheng began to develop and produce vascular additives, color reagents, chemiluminescent reagents and biological buffers in 2005. It has a long history of research on biological buffers, has a mature technology and R & D team, and has excellent sales service and logistics service. In addition to HEPES, there are Tris, caps, taps, bicine and pipes products for biological buffers. If you need to call, Desheng welcomes your call.
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