How do you make HEPES buffer from HEPES sodium salt?

A buffer solution of HEPES can be prepared by several methods. The free acid can be added to water, then titrated with approximately one- half mole equivalent of sodium hydroxide or potassium hydroxide to the precise pH desired, with adjustments made for final temperature and volume.

What is HEPES solution?

Overview. HEPES Buffer Solution is a biological buffer used in cell culture media. Selection of suitable nutrient medium is dependent on type of cell, conditions of culture, and degree of chemical definition required for the cell culture application.

How do you make a HEPES buffer?

For a 2x solution: Dissolve 0.8 g of NaCl, 0.027 g of Na2HPO4•2H2O, and 1.2 g of HEPES in a total volume of 90 ml of distilled H2O. Adjust the pH to 7.05 with 0.5 n NaOH, and then adjust the volume to 100 ml with distilled H2O.

How do you make a 1M solution of HEPES?

1 M HEPES, pH = 7.0

  1. Add 119.15 g HEPES (free acid) to a suitable container and make up to 400ml with distilled water.
  2. Add solid NaOH a few pellets at a time while mixing until the pH is ~6.8.
  3. Add concentrated NaOH dropwise to achieve pH = 7.0.
  4. Add distilled water to a final volume of 500 ml.

What is HEPES salt?

HEPES Sodium Salt is a zwitterionic salt which is used in the preparation of a HEPES buffer solution. Its pH range of 9.5-11.5 makes an ideal buffer for pH maintenance. HEPES Sodium Salt interferes with the Folin protein assay. The tendency of this buffer to form radicals makes it unsuitable for redox studies.

Is HEPES an acid or a base?

HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic sulfonic acid buffering agent, thus it has an acidic sulfonic acid group (pKa ~3) and a weakly basic piperazine group (pKa ~7.5).

What is the purpose of HEPES?

Just like other types of chemical buffers, HEPES buffer maintains the ion concentration of a solution at a level that’s ideal for a certain purpose. The pH level is maintained in this manner. Specifically, HEPES buffer is used to maintain the ideal conditions for cell culture.

What is HEPES buffered saline?

General description. HEPES buffered saline is a tissue culture buffer, which is used to maintain the pH of cell culture. It is water soluble and atmosphere independent. HEPES destroys magnesium in sodium chloride solutions.

What is the pH of HEPES?

6.8-8.2
HEPES buffer is one of the Good’s zwitterionic buffer with a pH range of 6.8-8.2. It is commonly used in cell culture medium as its pH is well maintained with changes in carbon dioxide concentration. Prepare 800 mL of dH2O in a suitable container. Add 238.3 g of Hepes to the solution.

What is HEPES free acid?

HEPES, Free Acid is a zwitterionic buffer ideally suited for cell culture in a range of 10 to 25mM. HEPES has a buffering range from 6.8 to 8.2. HEPES is also a component of Tris-HEPES buffer for polyacrylamide electrophoresis of proteins.

What is the role of HEPES in buffer?

How Does HEPES Buffer Work? Just like other types of chemical buffers, HEPES buffer maintains the ion concentration of a solution at a level that’s ideal for a certain purpose. The pH level is maintained in this manner. Specifically, HEPES buffer is used to maintain the ideal conditions for cell culture.

Is HEPES acidic or basic?