HEPES (4-hydroxyethyl piperazine ethanesulfonic acid) buffer commonly used buffer systems

 

HEPES (4-hydroxyethyl piperazine ethanesulfonic acid) is a commonly used biological buffer, widely used in cell culture, molecular biology experiments, and other biochemical research fields. Its unique properties make it an ideal choice for many experiments. When using HEPES buffer, there are two common buffer systems, namely HEPES buffer solution and HEPES buffer salt solution.

 

1、 HEPES buffer

HEPES buffer is usually composed of HEPES and alkali (such as NaOH). The following are the steps for preparing HEPES buffer:

(1) Dissolve 119.15 grams of HEPES in 400 milliliters of distilled water to ensure complete dissolution.

(2) Gradually add 0.5-1M NaOH aqueous solution dropwise and adjust the pH of the solution to the desired range. It should be noted that the effective pH buffer range of HEPES is 6.8-8.2.

(3) Bring the solution to 500 milliliters and use distilled water to bring it to volume.

(4) HEPES buffer is commonly used in biological experiments in situations where stable pH values need to be maintained, such as cell culture and protein expression. Its advantage lies in its ability to work under neutral pH conditions without toxicity to biological samples.

 

2、 HEPES buffer salt solution

In addition to HEPES and alkali, HEPES buffer salt solution also contains other ions. The following are the steps to prepare a HEPES buffer salt solution:

(1) Weigh 6.5 grams of HEPES, 8.0 grams of NaCl, and 0.198 grams of Na2HPO4 · 7H2O, respectively.

(2) Gradually add 0.5M NaOH aqueous solution dropwise and adjust the pH of the solution to the desired range.

(3) Bring the solution to 500 milliliters and use distilled water to bring it to volume.

(4) HEPES buffer salt solution is usually used for experiments that require specific ion concentrations, such as the need for calcium and magnesium ions in cell cell adhesion experiments.

 

3、 2 × HEPES buffer salt solution

This is a relatively complex HEPES buffer system suitable for specific experimental needs. The following is Preparation 2 × Steps for HEPES buffer salt solution:

(1) Weigh 1.6 grams of NaCl, 0.074 grams of KCl, 0.027 grams of Na2HPO4.2H2O, 0.2 grams of glucan or dextran, and 1 gram of HEPES, respectively, and add them to 90 milliliters of distilled water to ensure dissolution.

(2) Gradually add 0.5M NaOH aqueous solution dropwise and adjust the pH of the solution to the desired range.

(3) Bring the solution to 100 milliliters and use distilled water to bring it to volume.

(4) 2 × HEPES buffer salt solutions are suitable for special experimental designs, such as maintaining specific ion concentrations in cell culture.

 

In summary, HEPES buffers play an important role in biological science research. According to the experimental needs, suitable buffer systems can be selected to ensure the accuracy and stability of the experiment. As a supplier of HEPES, Desheng not only has advanced production equipment and strict quality control system, ensuring that the provided HEPES products have high purity and stable performance. At the same time, whether in product selection, application guidance, or experimental design, Desheng's professional team is able to provide timely assistance and solutions to customers. If you have any related needs, please feel free to contact us for purchase!