Cell Culture Buffering: Stabilizes pH in mammalian cell culture media, especially for CO2-independent or fluctuating CO2 conditions.

Live-Cell Imaging: Maintains physiological pH during microscopy and time-lapse imaging outside the incubator.

Bacterial and Yeast Culture: Provides pH stability in microbial growth media for organisms such as Escherichia coli and Bacillus subtilis.

Protein Purification: Used in ion-exchange, size-exclusion, and affinity chromatography buffers to maintain near-neutral pH.

Enzyme Assays: Provides a stable pH environment for enzymatic activity and kinetic measurements near pH 6.5–7.5.

Transfection Buffers: Component of HEPES-buffered saline (HBS/HEBS) for calcium phosphate and other transfection methods.

Electroporation: Included in electroporation buffers to support cell viability and maintain pH during pulsing.

Protein Lysis and Storage: Incorporated into lysis, dialysis, and storage buffers to preserve protein structure and function.