Description GoldBio’s HEPES, Free Acid is a zwitterionic biological buffer widely used in cell culture, biochemical assays, molecular biology, and protein research applications.
HEPES is particularly valued for its ability to maintain physiological pH in the range of approximately 6.8 to 8.2 while exhibiting relatively low interference with many biochemical reactions and cellular processes. Unlike bicarbonate-based buffering systems, HEPES provides effective buffering capacity independent of carbon dioxide equilibrium, making it useful in open-air experimental systems and microscopy applications.
HEPES free acid can be mixed with HEPES sodium salt to achieve the desired pH. Alternatively, HEPES free acid can be titrated with NaOH or KOH to achieve the desired pH.
Common Applications
(Click each for more information)
Mammalian Cell Culture Buffering
Purpose : To maintain stable physiological pH in mammalian cell culture systems.
How It Works : HEPES resists pH fluctuations caused by cellular metabolism and environmental CO2 variation.
Applications : Cell culture media preparation, live-cell maintenance, and suspension cell culture studies.
Good, N. E., Winget, G. D., Winter, W., Connolly, T. N., Izawa, S., & Singh, R. M. M. (1966). Hydrogen ion buffers for biological research. Biochemistry , 5(2), 467–477.
Electrophysiology and Patch Clamp Solutions
Purpose : To maintain controlled pH conditions in electrophysiological recording buffers.
How It Works : HEPES stabilizes ionic solution pH during electrical recording experiments involving isolated cells or tissues.
Applications : Patch clamp studies, ion channel analysis, and neurophysiology research.
Hamill, O. P., Marty, A., Neher, E., Sakmann, B., & Sigworth, F. J. (1981). Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Archiv , 391(2), 85–100.
Protein Purification and Enzyme Assays
Purpose : To maintain stable pH conditions that preserve enzyme activity and protein structure.
How It Works : HEPES buffers biochemical solutions while minimizing interference with enzymatic reactions and protein interactions.
Applications : Protein purification, enzyme kinetics, and biochemical assay development.
Ferguson, W. J., Braunschweiger, K. I., Braunschweiger, W. R., et al. (1980). Hydrogen ion buffers for biological research. Analytical Biochemistry , 104(2), 300–310.
Preparation of Physiological and Diagnostic Buffers
Purpose : To formulate stable buffered solutions used in physiological and diagnostic applications.
How It Works : HEPES provides reproducible buffering near neutral pH with low chemical reactivity.
Applications : Diagnostic reagent preparation, physiological saline buffers, and analytical chemistry applications.
Good, N. E., & Izawa, S. (1972). Hydrogen ion buffers. Methods in Enzymology , 24, 53–68.
Key Benefits
Maintains stable buffering capacity near physiological pH.
Effective buffering independent of CO2 equilibrium systems.
Compatible with many enzymes, proteins, and mammalian cell culture applications.
Commonly used in microscopy, electrophysiology, and live-cell imaging studies.
Low biological reactivity and minimal interference with many biochemical assays.
Storage/Handling
Store at room temperature.
Product Specifications
ULTRA PURE GRADE
Formula: C8 H18 N2 O4 S
MW: 238.30 g/mol
Storage/Handling: Store at room temperature.
PubChem Chemical ID: 23831
Video: HEPES Sodium Salt Video Protocol
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