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H3375

Sigma-Aldrich

HEPES

≥99.5% (titration)

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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:
7365-45-9
Molecular Weight:
238.30
Beilstein:
883043
EC Number:
230-907-9
MDL number:
MFCD00006158
UNSPSC Code:
12161700
PubChem Substance ID:
24895572
NACRES:
NA.25

Quality Level

400

Assay

≥99.5% (titration)

form

crystalline powder

storage condition

dry at room temperature

technique(s)

cell culture | mammalian: suitable

color

white

pH

5.0-6.5 (25 °C, 238 g/L)

useful pH range

6.8-8.2

pKa (25 °C)

7.5

solubility

water: 500 mg/mL, clear, colorless

suitability

suitable for component for culture media
suitable for enzyme extraction

application(s)

diagnostic assay manufacturing
general analytical
life science and biopharma
pharmaceutical

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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General description

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, also known as 4-(2-Hydroxyethyl)piperazine-1-ethane-sulfonic acid, is a zwitterionic N-substituted aminosulfonic acid buffer. It is effective in the pH 6.8-8.2 range, with a pKa of 7.48 at 25°C. Recognized as one of the best all-purpose buffers, especially in cell biology, biochemical, and biological research, HEPES is a dipolar ionic buffer and one of the Good′s buffers that does not form significant complexes with most metal ions. This property makes it suitable as a non-coordinating buffer in solutions with metal ions.

Widely utilized in cell culture, HEPES excels at maintaining physiological pH despite fluctuations in carbon dioxide concentration produced by cellular respiration, a feature not as prominent in bicarbonate buffers. It serves as an ampholytic separator for creating pH gradients in isoelectric focusing and exhibits less interference with DNA-restriction enzyme reactions compared to buffers with fewer substituted amine groups, such as Tris. Beyond these applications, HEPES may find application in various biological and biochemical studies, including immunoprecipitation, cell lysis, and live cell imaging. Its versatility makes it an indispensable tool in diverse research applications.

Application

HEPES has been used:
  • As a component in Danieau medium, which is used to prevent melanin pigment formation post gastrulation of zebra-fish embryo
  • As a component of Hanks′ solution used in the degradation testing of Iron
  • As a component of HEPES medium in the transfection of cell culture with plasmids
  • Along with RPMI-1640, L-glutamine, FBS, Sodium pyruvate and β-mercaptoethanol for the culturing of rat insulinoma cells
  • As a supplement in TCM199 medium for washing fresh oocytes required for nuclear transfer studies

Features and Benefits

  • High purity product for biochemical and biological research
  • Suitable for Cell culture
  • Highly soluble in water with a useful pH range of 6.8 - 8.2 and pKa of 7.5 at 25 °C

Other Notes

Go to Redi-Dri™ Product Listing RDD002
Easily compare specifications for HEPES products with the HEPES specification table.
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Legal Information

Redi-Dri is a trademark of Sigma-Aldrich Co. LLC

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

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  4. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  5. What is the purpose of using HEPES in cell culture?

    HEPES provides supplemental buffering for media in pH range of 7.2 to 7.6. It has no nutritional value to cells.

  6. What is the concentration of HEPES used in cell culture?

    HEPES is typically used in cell culture in the range of 10-25 mM.

  7. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Caveolae are highly immobile plasma membrane microdomains, which are not involved in constitutive endocytic trafficking.
Thomsen P, et al.
Molecular Biology of the Cell, 13(1), 238-250 (2002)
Electroformed pure iron as a new biomaterial for degradable stents: In vitro degradation and preliminary cell viability studies.
Moravej M, et al.
Acta Biomaterialia, 6(5), 1843-1851 (2010)
Culturing INS-1 cells on CDPGYIGSR-, RGD-and fibronectin surfaces improves insulin secretion and cell proliferation.
Kuehn C, et al.
Acta Biomaterialia, 8(2), 619-626 (2012)
High-resolution in situ hybridization to whole-mount zebrafish embryos.
Thisse C and Thisse B
Nature Protocols, 3(1), 59-69 (2008)
Structural, metabolic and developmental evaluation of ovulated rabbit oocytes before and after cryopreservation by vitrification and slow freezing.
Salvetti P, et al.
Theriogenology, 74(5), 847-855 (2010)
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