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H4034

Sigma-Aldrich

HEPES

BioPerformance Certified, ≥99.5% (titration), suitable for cell culture

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Synonym(s):
4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)
Empirical Formula (Hill Notation):
C8H18N2O4S
CAS Number:
Molecular Weight:
238.30
Beilstein:
883043
EC Number:
MDL number:
PubChem Substance ID:
NACRES:
NA.25

grade

BioPerformance Certified

Quality Level

Assay

≥99.5% (titration)

form

crystalline powder

technique(s)

cell culture | mammalian: suitable

impurities

endotoxin and total aerobic microbial count, tested

useful pH range

6.8-8.2

pKa (25 °C)

7.5

cation traces

Fe: ≤5 ppm

absorption

≤0.05 at 260 in H2O at 0.1 M
≤0.05 at 280 in H2O at 0.1 M

application(s)

diagnostic assay manufacturing

foreign activity

DNase, RNase, NICKase, protease, none detected

SMILES string

OCCN1CCN(CC1)CCS(O)(=O)=O

InChI

1S/C8H18N2O4S/c11-7-5-9-1-3-10(4-2-9)6-8-15(12,13)14/h11H,1-8H2,(H,12,13,14)

InChI key

JKMHFZQWWAIEOD-UHFFFAOYSA-N

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1 of 4

This Item
H3375H6147H7523
HEPES BioPerformance Certified, ≥99.5% (titration), suitable for cell culture

Sigma-Aldrich

H4034

HEPES

Premium Grade
HEPES ≥99.5% (titration)

Sigma-Aldrich

H3375

HEPES

-
HEPES BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)

Sigma-Aldrich

H6147

HEPES

Premium Grade
HEPES BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)

Sigma-Aldrich

H7523

HEPES

Premium Grade
assay

≥99.5% (titration)

assay

≥99.5% (titration)

assay

≥99.5% (titration)

assay

≥99.5% (titration)

form

crystalline powder

form

crystalline powder

form

powder

form

powder

technique(s)

cell culture | mammalian: suitable

technique(s)

cell culture | mammalian: suitable

technique(s)

cell culture | embryo: suitable

technique(s)

-

impurities

endotoxin and total aerobic microbial count, tested

impurities

-

impurities

-

impurities

Insoluble matter, passes filter test

cation traces

Fe: ≤5 ppm

cation traces

-

cation traces

-

cation traces

Al: ≤0.0005%, As: ≤0.0001%, Ba: ≤0.0005%, Bi: ≤0.0005%, Ca: ≤0.001%, Cd: ≤0.0005%, Co: ≤0.0005%, Cr: ≤0.0005%, Cu: ≤0.0005%, Fe: ≤0.0005%, K: ≤0.005%, Li: ≤0.0005%, Mg: ≤0.0005%, Mn: ≤0.0005%, Mo: ≤0.0005%, Na: ≤0.005%, Ni: ≤0.0005%, Pb: ≤0.0005%, Sr: ≤0.0005%, Zn: ≤0.0005%

General description

HEPES buffer does not confer cytotoxic effects on cells and thus can be used in animal cell cultures.
HEPES has been described as one of the best all-purpose buffers available for biological research. At biological pH, the molecule is zwitterionic, and is effective as a buffer at pH 6.8 to 8.2. HEPES has been used in a wide variety of applications, including tissue culture. It is commonly used to buffer cell culture media in air. HEPES finds its usage in in vitro experiments on Mg.
HEPES, or 4-(2-hydroxyethyl)piperazine-1-ethane-sulfonic acid, is a zwitterionic N-substituted aminosulfonic acid buffer. It demonstrates outstanding buffering capacity in the pH range of 6.8 to 8.2, boasting a pKa of 7.48 at 25°C. Widely acknowledged as one of the most versatile buffers, HEPES is extensively utilized in cell biology, biochemical, and biological research. A notable attribute is its non-reactivity with metal ions. Unlike many buffers, HEPES does not form significant complexes with most metals, making it suitable for applications involving metal ions without affecting their activity. This characteristic enhances its utility as a "non-coordinating buffer." In cell culture, HEPES excels in maintaining a stable physiological pH, even amidst fluctuations in carbon dioxide concentration resulting from cellular respiration. This sets it apart from bicarbonate buffers (NaHCO3), which, though commonly used, are less effective in pH stability.

HEPES also proves advantageous in various biological and biochemical processes. Its ampholytic nature makes it suitable as a separator for creating pH gradients in isoelectric focusing, a technique often employed in protein separation and analysis. Moreover, HEPES exhibits minimal interference with DNA-restriction enzyme reactions compared to buffers with fewer substituted amine groups, such as Tris. This makes it a preferred choice for applications involving DNA manipulation and analysis. Beyond these specific applications, HEPES is valuable in numerous other biological and biochemical processes, including immunoprecipitation, cell lysis, and live cell imaging. Its versatility, coupled with exceptional pH buffering capacity and minimal interaction with other molecules, establishes HEPES as an indispensable tool across diverse research domains.

Application

HEPES has been used:
  • To supplement Dulbecco′s modified Eagle′s medium to culture and maintain cell lines
  • As a component of platelet suspension buffer
  • To supplement Hank′s basic salt solution, which is used to wash pancreatic tissue
  • As a component of wash buffer and blocking buffer in the purification and quantification of protein with enzyme-linked immunosorbent (ELISA) assay
  • For the adjustment and maintenance of pH of biological solutions
  • As a component of Hank′s balanced salt solution (HBSS) and dissociation medium to study neuronal development
  • For homogenization of tissue and in the preparation of cytosolic and nuclear extract from cells
  • As a component of keratinocyte and fibroblast culture medium

Features and Benefits

  • Tested to confirm low levels of heavy metal contamination, ensuring suitability for various applications
  • Highly soluble in water with a useful pH range of 6.8 - 8.2 and pKa of 7.5 at 25 °C
  • Negligible metal ion binding
  • Less toxic to cells than other buffers such as Tris and phosphate
  • Stable in a wide pH range
  • Tested for Endotoxins and Total Aerobic Microbial Count
  • Free from DNase, NICKase, RNase, Endonuclease, Exonuclease and Protease

Other Notes

Easily compare specifications for HEPES products with the HEPES specification table.

Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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The slice overlay assay: a versatile tool to study the influence of extracellular signals on neuronal development.
Polleux F and Ghosh A
Science's STKE : Signal Transduction Knowledge Environment, 136, pl9-pl9 (2002)
Three-Dimensional Human Tissue Models of Wounded Skin
Egles C, et al.
Methods in Molecular Biology, 585, 345-359 (2010)
Complex thermorheology of living cells.
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Airway epithelial cell PPAR? modulates cigarette smoke-induced chemokine expression and emphysema susceptibility in mice.
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Protocols

This protocol describes a method for chemical cross-linking of proteins using formaldehyde. With the exception of zero-length cross-linkers, formaldehyde has the shortest cross-linking span (~2-3 Å) of any cross-linking reagent, thus making it an ideal tool for detecting specific protein-protein interactions with great confidence.

To determine the molecular weights of protein antigens, to study protein/protein interactions, to determine specific enzymatic activity, to monitor protein post-translational modifications and to determine the presence and quantity of proteins.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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