- Prepare 800 mL of dH2O in a suitable container.
- Add 238.3 g of HEPES to the solution.
- Adjust solution to desired pH by 10N NaOH.
- Add dH2O until the volume is 1.
How do you dilute HEPES?
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.21 Dec 2013
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.16 May 2017
How do you make HEPES buffered saline?
2.1. HEPES-buffered saline (HBS) solution: Prepare 2 × HBS stock by adding the following to 400 ml dH2O: 50 ml 1 M HEPES, 28 ml 5 M NaCl and 1.5 ml 0.5 M Na2HPO4. After adjusting the pH to 7.05–7.10 with 5 M and 1 M NaOH, bring the total volume to 500 ml with dH2O.
What does HEPES do in cell culture media?
The most commonly used concentration is 25mM. HEPES has no nutritional benefit to cells. It is added to the media solely for extra buffering capacity when cell culture requires extended periods of manipulation outside of a CO2 incubator.
Is HEPES necessary?
HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) is a commonly used zwitterionic buffer with a pKa of 7.3 at 37 °C. HEPES should be used in addition to, not instead of, sodium bicarbonate, as it is important to maintain sufficient bicarbonate in the medium for nutritional purposes.
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).9 Sept 2020
Is HEPES an acid?
HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic sulfonic acid buffering agent; one of the twenty Good's buffers.
What is the pH of 1M HEPES?
7.0
What is the function of HEPES?
HEPES : - A biological buffer (exist in a zwitterionic form ). It maintains physiological pH in cell culture when compared to bicarbonate buffers despite the changes in carbon dioxide concentration.
Why is HEPES added to cell culture media?
HEPES has no nutritional benefit to cells. It is added to the media solely for extra buffering capacity when cell culture requires extended periods of manipulation outside of a CO2 incubator.
How does HEPES work as a buffer in a cell culture medium?
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.14 Jul 2021