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About HEPES - HEPES is a short name for 2-(4-(2-hydroxyethyl)-1-piperazinyl)-ethanesulfonic acid, a buffer substance. - Chemical properties - HEPES is used for biochemical, molecular biological and microbiological purposes as a buffer substance from the group of good buffers. With an acid constant of pKs = 7.55 (at 20 °C), it has a good buffering capacity between pH 6.8 and 8.2. The temperature dependence is ΔpKs/ΔT = -0.014 K-1.
HEPES is a widely used buffer in biological studies. There is only one restriction in the use of this buffer, because it interferes with the Folin protein assay. In cell culture media, it is employed as a substitute for the bicarbonate buffer at a concentration of 25 mM or as a supplement to the bicarbonate buffer (concentration 10 - 15 mM).The addition of 20 mM HEPES to TBE buffer improves the migration behaviour of certain samples in the SSCP analysis (single-strand conformation polymorphism), with a higher resolution especially if small differences in the sequences between certain bands are wanted (5).
"HEPES (or 4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid) is a zwitterionic organic compound commonly used as a buffer solution in biochemistry. It belongs to the Good buffers. Since Good's work in 1966, it has been widely preferred to bicarbonate buffer for cell culture because of its better stability at physiological pH values.
HEPES is among the buffers with the best properties for biological studies (1,3,4). In cell culture, HEPES is used as a replacement for the bicarbonate buffer (then often at a concentration of 25 mM) or as an additional buffer to the bicarbonate (then often at a concentration of 10 - 15 mM). HEPES-buffered media are used especially when the stability of the pH value in the medium plays an important role. pH-value-sensitive cell culture systems receive additional buffering in the range of pH 7.2 - 7.6 through the addition of HEPES. Fluctuations in the pH value occur naturally through the metabolism of the cultured cells, but also with changes in the CO₂ concentration in the incubator or the surrounding environment (2). HEPES experiences limitations in usability in that it cannot be used when protein determination is to be performed after Folin. In addition, the piperazine ring system forms radicals under certain conditions. This buffer is therefore not suitable for the study of redox processes in biochemistry (5).The addition of 20 mM HEPES to the TBE buffer, improves the migration behavior of certain samples in SSCP (single-strand conformation polymorphism) analysis, where the resolution of particularly small sequence differences between individual bands is important (6)."
FAQs
What is HEPES?
HEPES is the abbreviated name for 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, formula C8H18N2O4S. It is an organic compound also called 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). It is widely used in biochemistry and molecular biology as a buffering agent to maintain physiological pH. It is a zwitterionic buffer (it tends to donate or accept protons, H+ ions, depending on the pH of the aqueous solution in which it is found), derived from piperazine, classified as a Good's buffer. It has a pKa of 7.48 at 25 °C, with effective buffering capacity in a pH range from 6.8 to 8.2. What is 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid?
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, formula C8H18N2O4S, is an organic compound commonly referred to as HEPES, other names being 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). It is widely used in biochemistry and molecular biology as a buffering agent to maintain physiological pH. It is a zwitterionic buffer (it tends to donate or accept protons, H+ ions, depending on the pH of the aqueous solution in which it is found), derived from piperazine, classified as a Good's buffer. It has a pKa of 7.48 at 25 °C, with effective buffering capacity in a pH range from 6.8 to 8.2. What is 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid?
4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, formula C8H18N2O4S, is an organic compound commonly referred to as HEPES, other names being 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). It is widely used in biochemistry and molecular biology as a buffering agent to maintain physiological pH. It is a zwitterionic buffer (it tends to donate or accept protons, H+ ions, depending on the pH of the aqueous solution in which it is found), derived from piperazine, classified as a Good's buffer. It has a pKa of 7.48 at 25 °C, with effective buffering capacity in a pH range from 6.8 to 8.2. What is 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid?
2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, formula C8H18N2O4S, is the IUPAC name for the organic compound commonly referred to as HEPES, other names being 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid or N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid). It is widely used in biochemistry and molecular biology as a buffering agent to maintain physiological pH. It is a zwitterionic buffer (it tends to donate or accept protons, H+ ions, depending on the pH of the aqueous solution in which it is found), derived from piperazine, classified as a Good's buffer. It has a pKa of 7.48 at 25 °C, with effective buffering capacity in a pH range from 6.8 to 8.2. What is N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid)?
N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid), formula C8H18N2O4S, is an organic compound commonly referred to as HEPES, other names being 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). It is widely used in biochemistry and molecular biology as a buffering agent to maintain physiological pH. It is a zwitterionic buffer (it tends to donate or accept protons, H+ ions, depending on the pH of the aqueous solution in which it is found), derived from piperazine, classified as a Good's buffer. It has a pKa of 7.48 at 25 °C, with effective buffering capacity in a pH range from 6.8 to 8.2. What does HEPES mean for?
HEPES is the abbreviated name for 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, formula C8H18N2O4S. It is an organic compound also called 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). It is widely used in biochemistry and molecular biology as a buffering agent to maintain physiological pH. It is a zwitterionic buffer (it tends to donate or accept protons, H+ ions, depending on the pH of the aqueous solution in which it is found), derived from piperazine, classified as a Good's buffer. It has a pKa of 7.48 at 25 °C, with effective buffering capacity in a pH range from 6.8 to 8.2. How is HEPES used?
HEPES is widely used for buffering biological samples because of its buffering capacity at physiological pH and its lower toxicity compared to other similar compounds; it is also used as a component of cell cultures, e.g. in the pharmaceutical industry. In cell culture media, it is used as a substitute for bicarbonate buffer at a concentration of 25 mM or as a supplement to bicarbonate buffer (concentration 10 - 15 mM). The addition of 20 mM HEPES to TBE buffer improves the migration behavior of certain samples in SSCP (single strand conformation polymorphism) analysis, with higher resolution, especially if small sequence differences between certain bands are desired. What is HEPES used for?
HEPES is widely used for buffering biological samples because of its buffering capacity at physiological pH and its lower toxicity compared to other similar compounds; it is also used as a component of cell cultures, e.g. in the pharmaceutical industry. In cell culture media, it is used as a substitute for bicarbonate buffer at a concentration of 25 mM or as a supplement to bicarbonate buffer (concentration 10 - 15 mM). The addition of 20 mM HEPES to TBE buffer improves the migration behavior of certain samples in SSCP (single strand conformation polymorphism) analysis, with higher resolution, especially if small sequence differences between certain bands are desired. What are the applications of HEPES?
HEPES is widely used for buffering biological samples because of its buffering capacity at physiological pH and its lower toxicity compared to other similar compounds; it is also used as a component of cell cultures, e.g. in the pharmaceutical industry. In cell culture media, it is used as a substitute for bicarbonate buffer at a concentration of 25 mM or as a supplement to bicarbonate buffer (concentration 10 - 15 mM). The addition of 20 mM HEPES to TBE buffer improves the migration behavior of certain samples in SSCP (single strand conformation polymorphism) analysis, with higher resolution, especially if small sequence differences between certain bands are desired. What is the CAS number of HEPES?
The CAS number of HEPES is 7365-45-9. What is the CAS number 7365-45-9?
The CAS number is a unique numeric identifier, designating a single, specific substance. The CAS number 7365-45-9 corresponds to HEPES, formula C8H18N2O4S. It is an organic compound also called 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). To which substance does the CAS number 7365-45-9 correspond?
The CAS number is a unique numeric identifier, designating a single, specific substance. The CAS number 7365-45-9 corresponds to HEPES, formula C8H18N2O4S. It is an organic compound also called 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) or 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (IUPAC name). What is the pKa of HEPES?
The pKa of HEPES is 7.48 at 25 °C. What is the effective pH range of HEPES?
HEPES has an effective pH range of 6.8 to 8.2. What is the molecular formula of HEPES?
The molecular formula of HEPES is C8H18N2O4S. What is the molar mass of HEPES?
The molar mass of HEPES is 238.31 g/mol. What is the molecular weight of HEPES?
The molecular weight of HEPES is 238.31 g/mol. How to prepare a 1 mol/L (1 M) HEPES buffer?
To prepare 1 L of 1 mol/L (1 M) HEPES buffer solution, dissolve 238.3 g of HEPES in 800 mL of distilled water, adjust the pH with a pH meter to the desired value with 10 mol/L (10 N) NaOH and add distilled water up to 1 L. What is the pH of HEPES?
The pH of a 1% aqueous solution of HEPES is between 4.7 and 6.0. What is the melting point of HEPES?
The melting point of HEPES is between 234 and 238°C. What is the solubility of HEPES?
The solubility of HEPES in water is 400 g/L at 20 °C. What is the difference between HEPES, HEPPS and HEPPSO?
Chemically, HEPES, HEPPS and HEPPSO are all alkanesulfonic acids derived from piperazine. All three are zwitterionic buffers, classified as Good buffers. The difference is in the carbon chain length between the amino group and the sulfonic acid, therefore the pKa (and effective pH range) are different. Where to buy HEPES?
You can purchase HEPES from ITW Reagents through its worldwide network of distributors, or through the online store if you are a registered ITW Reagents customer. If you wish to purchase HEPES for production processes, please contact our distributors. Follow this link to find a distributor in your country https://www.itwreagents.com/rest-of-world/en/distributors-rw.
Literature
(1) Good, N.E. et al. (1966) Biochemistry 5, 467-477. Synthesis and characterization of various 'H+ buffers' for biological research. (2) Shipman, C. (1969) Proc. Soc. Exp. Biol. Med. 130, 305-310. HEPES as a buffer in cell culture. (3) Good, N.E. & Izawa, S. (1972) Methods Enzymol. 24, 53-68. Further discussion on the selection of buffers in biological research, here with emphasis on their suitability for the study of photosynthetic processes. (4) Ferguson, W.J. et al. (1980) Anal. Biochem. 104, 300-310. Buffers to buffer hydrogen ions for biological research: synthesis of other buffers and test in cell culture, plant pathology, and virology. (5) Grady, J.K. et al. (1988) Anal. Biochem. 173, 111-115. Radicals from Good buffers. (6) Liu, Q. & Sommer, S.S. (1998) Biotechniques 25, 50-56. The SSCP phenomenon: addition of HEPES affects electrophoretic mobility.
