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HEPES solution

1 M, pH 7.0-7.6, sterile-filtered, BioReagent, suitable for cell culture

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N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)
CAS Number:
MDL number:
PubChem Substance ID:



Quality Level

product line




storage condition

(Tightly closed. Dry)


1 M


cell culture | mammalian: suitable


endotoxin, tested



useful pH range



suitable for component for culture media
suitable for electrophoresis


cell analysis
life science and biopharma
sample preparation

storage temp.


SMILES string




InChI key


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General description

HEPES Buffer Solution, a sterile-filtered 1M solution of 4-(2-hydroxyethyl)piperazine-1-ethane-sulfonic acid, serves as a vital biological buffer in cell biology, biochemical, and biological research. This zwitterionic buffer enjoys widespread adoption due to its effective pH range of 6.8 to 8.2, making it suitable for a diverse array of research applications. HEPES excels as a buffering agent in cell culture media, offering several advantages over alternatives like Tris and phosphate. Notably, HEPES exhibits lower cell toxicity, ensuring optimal cell health and viability during experiments.

Furthermore, it surpasses bicarbonate buffers (NaHCO3) in its ability to maintain physiological pH despite fluctuations in carbon dioxide concentration arising from cellular respiration, a key factor in maintaining cellular homeostasis. Beyond its exceptional performance in cell culture, HEPES may also find application in various biological and biochemical studies. It may also be valuable in techniques like immunoprecipitation, cell lysis, and live cell imaging, providing a stable and controlled environment for experimental procedures. The combination of its versatility, exceptional pH buffering capacity, and minimal interaction with other molecules positions HEPES as an indispensable tool across diverse research fields.
It is typically used in cell culture at concentration between 5mM to 30 mM. 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 has been used as a component of:
  • Hank’s balanced salt solution, Dulbecco’s modified eagle’s medium and no-glucose DMEM, which are used for the preparation of tissue slices
  • Buffers used in localized conductance measurements on the basolateral side of culture inserts to examine cation selectivity
  • cell culture media to provide buffer capacity to maintain the pH of the cell line
  • buffer in decellularization of porcine nucleus pulposus

Features and Benefits

  • High purity product for biochemical and biological research
  • Sterile filtered and cell culture tested
  • Ready Solution reduces the need
  • Useful for the pH range of 6.8 - 8.2 and pKa of 7.5 at 25 °C
  • Ready available solution reduce the need for preparation time


Concentrations of HEPES over 40mM may be toxic to some cell types.

Preparation Note

Prepared from HEPES free acid and cell culture grade water.

Other Notes

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Storage Class Code

10 - Combustible liquids



Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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Kaixi Tang et al.
PLoS computational biology, 14(7), e1006303-e1006303 (2018-07-06)
Focal adhesions are protein complexes that anchor cells to the extracellular matrix. During migration, the growth and disassembly of these structures are spatiotemporally regulated, with new adhesions forming at the leading edge of the cell and mature adhesions disassembling at
Assessing the corrosion of biodegradable magnesium implants: a critical review of current methodologies and their limitations.
Kirkland Nt
Acta Biomaterialia, 8, 925- 936 (2012)
Chiao-Chen Chen et al.
Analytical chemistry, 85(7), 3621-3628 (2013-02-21)
Elucidation of epithelial transport across transcellular or paracellular pathways promises to advance the present understanding of ion transport and enables regulation of cell junctions critical to the cell and molecular biology of the epithelium. Here, we demonstrate a new instrumental
Zuzana Keckesova et al.
Nature, 543(7647), 681-686 (2017-03-23)
Post-mitotic, differentiated cells exhibit a variety of characteristics that contrast with those of actively growing neoplastic cells, such as the expression of cell-cycle inhibitors and differentiation factors. We hypothesized that the gene expression profiles of these differentiated cells could reveal
Agnieszka Pozarska et al.
American journal of physiology. Lung cellular and molecular physiology, 312(6), L882-L895 (2017-03-21)
Postnatal lung maturation generates a large number of small alveoli, with concomitant thinning of alveolar septal walls, generating a large gas exchange surface area but minimizing the distance traversed by the gases. This demand for a large and thin gas


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