12 uses of HEPES Buffer you didn’t know

forward from: http://www.hopaxfc.com

Hepes is a zwitterionic biological buffer often used in biological and biochemical research. It was one of the first Good’s buffers described by Norman E. Good in 1966. Although it is an extremely popular buffering agent in labs all over the world, what is Hepes actually suitable for?

We collected reliable information from renowned academic sources in order to help scientists understanding better Hepes’ characteristics and suitability for different biotechnology applications (cell culture, chromatography, and others). Check out our findings!

Hepes basic information:

• CAS Number: 7365–45–9
• Molecular Weight: 238.3
• Formula: C8H18N2O4S
• Useful ph range: 6.8–8.2
• pKa (25°C): 7.45–7.65
• Specifications / Price: Click here

What is Hepes recommended for?

For most studies with metal ions(1)

1. As a good substitute of Tris and phosphate in studies with metal ions(2)
2. For environmental, analytical and biological studies(3)
3. As a binding buffer and an eluent during cation exchange chromatography(4)
4. As a grinding buffer in plant studies(5)
5. As a running buffer in gel electrophoresis(6)
6. As a buffer in electroporation(7)
7. To buffer mammalian cell cultures(8)
8. As buffer for in vitro fertilization and embryo culture(9)
9. For Bradford or bicinchoninic acid (BCA) assays
10. For researches regarding Amphidinium carterae, since it is not toxic to this algae(10)
11. It has been introduced in the extraction buffer to prevent the damage of proteins in red blood cells(3)

Which concerns should you have before choosing Hepes for your research?

• It affects membrane potentials in neuronal cells(11)
• It interferes with Lowry protein determination(12)
• It is not suitable for redox studies(3)
• It is toxic to small crustaceans (water fleas), such as Daphnia magna and Daphnia pulex(13)
• It interferes with phenolic oxidation by peroxidases(14)
• It can affect iron autoxidation rates(15)
• It is not recommended when a protein assay is performed by using Folin reagent

Useful tips about Hepes:

• It is very soluble in water
• It has negligible metal ion binding
• It inactivates DEPC, a popular RNase inhibitor

Hopax Hepes Buffer

Hopax Fine Chemicals is among the largest manufacturers of Hepes in the world. Our products are shipped daily to top research centers and biotech companies in Europe, America and Asia.

What we offer:

• Product straight from our manufacturing sites
• Worldwide shipping to your door
• Assistance with shipping
• Small and bulk packages (from grams to tons)
• International quality standards
• After-sales service with English speaking staff

References:

1 Good, N.E. & Izawa, S. (1972) Methods Enzymol. 24, 53–68. Available at https://www.sciencedirect.com/science/article/pii/007668797224054X?via%3Dihub

2 Brom, M., Joosten, L., Oyen, W. J., Gotthardt, M. & Boerman, O. C. (2012) EJNMMI Res., 2, 4. Available at https://www.ncbi.nlm.nih.gov/pubmed/22284727

3 Ferreira C. M., Pinto I.S., Soares, E.V., & Soares H.M. (2015) (Un)suitability of the use of pH buffers in biological, biochemical and environmental studies and their interaction with metal ions — a review, Royal Society of Chemistry 30989–31003. Available at https://repositorium.sdum.uminho.pt/bitstream/1822/38712/1/document_19948_1.pdf

3 Grady, J. K., Chasteen, N. D., & Harris, D. C. (1988). Radicals from “Good’s” buffers. Analytical Biochemistry, 173(1), 111–115. Available at https://www.ncbi.nlm.nih.gov/pubmed/2847586

4 Susin, S. A., Lorenzo, H. K., Zamzami, N., Marzo, I., Brenner, C., Larochette, N., & Kroemer, G. (1999). Mitochondrial release of caspase-2 and-9 during the apoptotic process. The Journal of Experimental Medicine, 189(2), 381–394. Available at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2192979/

5 Soltis, D. E., Haufler, C. H., Darrow, D. C., & Gastony, G. J. (1983). Starch gel electrophoresis of ferns: a compilation of grinding buffers, gel and electrode buffers, and staining schedules. American Fern Journal, 9–27. Available https://www.jstor.org/stable/pdf/1546611.pdf

6 Michels, D. A., Hu, S., Schoenherr, R. M., Eggertson, M. J., & Dovichi, N. J. (2002). Fully automated twodimensional capillary electrophoresis for high sensitivity protein analysis. Molecular & Cellular Proteomics, 1(1), 69–74. Available at https://www.ncbi.nlm.nih.gov/pubmed/12096142

7 Fromm, M., Taylor, L. P., & Walbot, V. (1985). Expression of genes transferred into monocot and dicot plant cells by electroporation. Proceedings of the National Academy of Sciences, 82(17), 5824–5828. Available at https://www.jstor.org/stable/26200

8 Itagaki, A., & Kimura, G. (1974). TES and HEPES buffers in mammalian cell cultures and viral studies: problem of carbon dioxide requirement. Experimental Cell Research, 83(2), 351–361. Available at https://www.sciencedirect.com/science/article/pii/0014482774903498

9 Mahadevan, M. M., Fleetham, J., Church, R. B., & Taylor, P. J. (1986). Growth of mouse embryos in bicarbonate media buffered by carbon dioxide, hepes, or phosphate. Journal of In Vitro Fertilization and Embryo Transfer, 3(5), 304–308. Available at https://link.springer.com/article/10.1007%2FBF01133390

10 Vasconcelos, M., Azenha, M., & Lage O. (1996) Electrochemical Evidence of Surfactant Activity of the Hepes pH Buffer Which May Have Implications on Trace Metal Availability to Cultures in Vitro. Analytical Biochemistry 241, 248–253. Available at https://www.ncbi.nlm.nih.gov/pubmed/8921194

11Cowan, A. I., & Martin, R. L. (1996) Brain Res., 717, 69–75. Available at https://www.sciencedirect.com/science/article/pii/0006899396000522

12 Peterson, G. L. (1979). Review of the Folin phenol protein quantitation method of Lowry, Rosebrough, Farr and Randall. Analytical Biochemistry, 100(2), 201–220. Available at https://www.sciencedirect.com/science/article/pii/0003269779902227

13 Keating, K. I., Caffrey, P. B., & Dagbusan, B. C. (1996) Environ. Toxicol. Chem., 15, 348–352. Available at http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.535.9527&rep=rep1&type=pdf

14 Baker, C. J., Mock, N. M., Roberts, D. P., Deahl, K. L., Hapeman C. J., Schmidt W. F. & Kochansky J. (2007) Free Radical Biol. Med., 43, 1322–1327. Available at http://europepmc.org/abstract/med/17893045

15 Yang, X. & Chasteen, N. D. (1999) Biochem. J., 618, 615–618. Available at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1220094/