HEPES

HEPES, or 4-(2-hydroxyethyl)piperazine-1-ethane-sulfonic acid, serves as a zwitterionic N-substituted aminosulfonic acid buffer with exceptional buffering capacity (pH 6.8-8.2, pKa 7.48 at 25°C). Recognized for versatility, it is extensively employed in cell biology, biochemical, and biological research. Its noteworthy feature lies in its neutrality towards metal ions, making it a preferred "non-coordinating buffer" suitable for solutions containing metals without affecting their activity. In cell biology studies, HEPES excels at maintaining stable physiological pH despite carbon dioxide fluctuations. It outperforms bicarbonate buffers commonly used in this context.

HEPES extends its utility as a preferred separator in creating pH gradients for isoelectric focusing, a technique in protein separation. Beyond these applications, it proves beneficial in immunoprecipitation, cell lysis, live cell imaging, establishing its indispensability across diverse research areas due to its versatility and minimal interaction with other molecules.

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

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