How do you make a sodium HEPES buffer?

How do you make a sodium HEPES buffer?

HEPES Buffer (1 M, 7.5 pH) Preparation and Recipe

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 238.3 g of HEPES to the solution.
3. Adjust solution to desired pH by 10N NaOH.
4. Add dH2O until volume is 1 L.

How do you make a 1M HEPES buffer?

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 sodium salt?

HEPES sodium salt is a buffering agent used in biochemistry and molecular biology that was selected and described by Good et al. It is a zwitterionic, piperazinic buffer that is useful for a pH range of 6.8 – 8.2. This buffer is capable of forming radicals and is therefore not suitable for redox reactions.

How do you make 20mm HEPES?

The method calls for 20 mM HEPES pH 7.2. You can prepare this in a number of ways. One typical way is prepare a stock solution of 0.5 or 1 M HEPES from solid, with the pH adjusted to 7.2 using NaOH. This is then diluted to make the 20 mM buffer.

How do you make a HEPES lysis buffer?

For 500 ml of lysis/IP buffer, combine 25 ml of 1M HEPES/KOH (pH7. 5), 14 ml of 5 M NaCl, 1 ml of 500 mM EDTA, and 50 ml of 10% Triton X-100 in 400 ml H2O. Add 0.5 g of sodium deoxycholate and make up the volume to 500 ml with H2O.

What do you dilute HEPES in?

Add 2.38 g of HEPES to an appropriate beaker (100-200 mL beaker in this case). Add about 80 mL of deionized water to the beaker. Add a stir bar to the beaker and leave it on a stir plate until completely dissolved (~1 min). Begin monitoring pH of the solution.

Is HEPES better than Tris?

The pKa of HEPES is 7.5 and the pKa of Tris is 8.1. If you want to maintain the pH at 7.5, HEPES is better than Tris. Because HEPES is a sulfonic acid, you have to add a base to neutralize it, so the solution ends up with a cation, usually Na+.

What is in HEPES buffer?

This buffer is classified as a zwitterionic sulphonic acid buffering agent with the IUPAC chemical name 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid. Its chemical formula can be written as C8H18N2O4S, while its structural formula is written as: HEPES buffer has the following properties: Molar mass: 238.3012 g/mol.

Can I autoclave HEPES buffer?

About the sterilization method of HEPES, HEPES powder is resistant to high temperatures with its melting point reaching 200℃, thus, it will not get degraded by autoclaving, or you can filter HEPES Buffer solutions by a 0.2-micrometer syringe filter.

Can you autoclave HEPES buffer?

4 Answers. From here: Some buffers (e.g., MOPS and HEPES) cannot be autoclaved because they degrade upon heating.

How do you adjust the pH of HEPES?

Procedure

1. Add 2.38 g of HEPES to an appropriate beaker (100-200 mL beaker in this case).
2. Add about 80 mL of deionized water to the beaker.
3. Add a stir bar to the beaker and leave it on a stir plate until completely dissolved (~1 min).
4. Begin monitoring pH of the solution.
5. Add one NaOH pellet to raise the pH towards 7.4.

When would you use a HEPES buffer?

HEPES is widely used in cell culture, largely because it is better at maintaining physiological pH despite changes in carbon dioxide concentration (produced by aerobic respiration) when compared to bicarbonate buffers, which are also commonly used in cell culture.