How to prepare hepes buffer

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.

How to make pipes and precautions

Pipes is a member of piperazine buffer family, and belongs to the same family as HEPES. It is a good's buffer for amphoteric ions, and it is effective in the pH range of 6.1 to 7.5. Because the pH range is slightly different from that of HEPES, the concentration of pipe is only half of that of HEPES. The buffer does not complex with metals, so it can be used in solutions containing metal ions. It can also be used for chromatographic analysis, immobilization of plant cells by electron microscopy, and as a substitute for glyphosate buffer.

Preparation steps of pipes:

To prepare 1 litre of 1m pipes free acid buffer solution, 302.37g   Pipes free acid added to 600 ml DH   two   O. Adjust to the desired pH with 10N sodium hydroxide. You can use a table in the 1m pipes PDF protocol. Using DH   two   O fill to the final volume of 1L and sterilize with a filter or autoclave. Stored in 4 ? C。 If you are using sodium pipes, use 1m pipes Na PDF protocol instead.

matters needing attention:

The free acid of pipes is easy to dissolve until the pH value of the solution rises to 6.5, but the sodium salt of pipes is easy to dissolve. If buffer solution in the form of free acid is used, use sodium hydroxide to convert it into sodium salt to improve solubility at lower pH. Like HEPES, pipes is not suitable for experiments involving redox reactions, as it can lead to the formation of free radicals.