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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/REAXYS Number:
883043
EC Number:
MDL number:
PubChem Substance ID:
NACRES:
NA.25

grade

BioPerformance Certified

Quality Level

assay

≥99.5% (titration)

form

crystalline powder

storage condition

dry at room temperature

technique(s)

cell culture | mammalian: suitable
competitive inhibition ELISA: suitable

impurities

endotoxin and total aerobic microbial count, tested

color

white

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

suitability

suitable for Western blot
suitable for component for culture media

application(s)

clinical research
diagnostic assay manufacturing
general analytical
life science and biopharma

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
H7523H3375H23830
HEPES BioPerformance Certified, ≥99.5% (titration), suitable for cell culture

H4034

HEPES

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

H7523

HEPES

Premium Grade
HEPES ≥99.5% (titration)

H3375

HEPES

-
HEPES reagent grade, ≥99% (titration), crystalline

H23830

HEPES

Essential Grade
assay

≥99.5% (titration)

assay

≥99.5% (titration)

assay

≥99.5% (titration)

assay

≥99% (titration)

grade

BioPerformance Certified

grade

-

grade

-

grade

reagent grade

Quality Level

400

Quality Level

400

Quality Level

400

Quality Level

200

form

crystalline powder

form

powder

form

crystalline powder

form

crystalline

application(s)

clinical research
diagnostic assay manufacturing
general analytical
life science and biopharma

application(s)

diagnostic assay manufacturing

application(s)

diagnostic assay manufacturing
general analytical
life science and biopharma
pharmaceutical

application(s)

-

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

For additional information on our range of Biochemicals, please complete this form.

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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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)
Blood compatibility of surfaces with superlow protein adsorption.
Zhang Z, et al.
Biomaterials, 29(32), 4285-4291 (2008)
Airway epithelial cell PPAR? modulates cigarette smoke-induced chemokine expression and emphysema susceptibility in mice.
Solleti SK et al.
American Journal of Physiology. Lung Cellular and Molecular Physiology, 309, L293-L293 (2015)
Use of a new buffer in the culture of animal cells.
Williamson JD and Cox P
The Journal of General Virology, 2, 309-309 (1968)

Protocols

Formaldehyde cross-linking protocol detects specific protein interactions with confidence.

Techniques for protein antigen molecular weight determination, protein interactions, enzymatic activity, and post-translational modifications.

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