How to prepare biological buffer HEPES

Details:

Many biological experiments need to use buffer to maintain effective pH, which is very important, because proteins and enzymes are sensitive to pH changes, so it is very important to choose the right buffer for current and downstream experiments. In this article, I will briefly discuss some important factors to be considered in the configuration and use of HEPES and pipes, hoping to be helpful to you.

How to make HEPES and precautions

HEPES is a zwitterionic good's buffer, effective in the pH range of 6.8 to 8.2. This makes HEPES an effective buffer at physiological pH. The buffer can be used in cell culture medium of many kinds of organisms. It can also be used as a binding buffer in protein research, used in cation exchange elution experiments, and as a running buffer in gel electrophoresis.

The preparation steps of HEPES are as follows

To prepare 1 liter of 1m HEPES buffer, add 238.30 G   Dissolution of goldbio HEPES   At 750 mL DH   two   O. Adjust to the desired pH with 10N sodium hydroxide. You can use a table in 1m HEPES PDF protocol. Using DH   two   O fill to final volume 1L and sterilize through filter or autoclave. The buffer was stored at 4 ℃. If you are using HEPES sodium salt, please use HEPES Na solution instead.

matters needing attention:

HEPES is water-soluble, inexpensive, and generally bioinert. However, it does participate in the redox reaction of producing free radicals and interferes with the reaction between DNA and restriction endonuclease, but due to steric hindrance, its interference degree is less than that of Tris. It is also important to note that when using heterojunction protein channels, HEPES should not be used because it inhibits its function. For toxicity studies, choose HEPES sodium salt instead of free acid.