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Why is HEPES the top buffer?

Now there is a saying that HEPES is the top organic chemical buffer. As we all know, there are many buffers, such as hydrochloric acid-sodium citrate, citric acid-sodium citrate, and acetic acid-sodium acetate, etc., that we know. For these buffers, what exactly is the advantage of HEPES?

Disadvantages of most buffers on the market

Most buffer solvents have one problem or another. For example, when the carbon dioxide level changes, it will affect the buffer's action. Some buffers can negatively affect the metal substrate when protein deposition is observed by electron microscopy. Some buffers tend to bind to calcium, so they cannot be used in the presence of calcium ions. So how to solve these problems? So, we are going to introduce HEPES as a buffer here.

The action range of HEPES

HEPES is also considered to be one of the top general-purpose buffers used in biological research. It is amphoteric in the presence of biological pH and is effective as a buffer between pH 6.8 and 8.2. HEPES is used in many different applications, such as tissue culture.

Some advantages of HEPES

Firstly, since cell metabolism and growth in culture may result in dramatic changes in medium pH, the addition of a sufficient amount of biochemical buffer to the medium of the culture is necessary to create an environment where cells can grow in a controlled manner. The ideal culture conditions for most cells are pH 7.2 - pH 7.4, and HEPES is well suited for cell culture with a buffer range right between pH 6.8 - pH 8.2.

Some considerations for HEPES

Finally, there are some caveats when we use HEPES. HEPES exposed to light sources produce hydrogen peroxide, which is not a problem in bicarbonate-based cell culture buffers. Therefore, it is highly recommended that solutions containing HEPES be stored under dark conditions whenever possible to avoid combustion.

September 2022