Mammalian Cell Culture: Stabilizes extracellular pH (CO2-independent or low-CO2 conditions) in media and supplements for sensitive cells.

Live-Cell Imaging: Maintains physiological pH during microscopy and time-lapse experiments outside incubators.

Protein Purification: Buffers affinity, ion-exchange, and size-exclusion chromatography at pH 7.5 to preserve protein stability and activity.

Enzyme Assays: Provides a stable, low-temperature- and salt-sensitive pH environment for accurate kinetic measurements.

Nucleic Acid Workflows: Buffers reactions and sample prep where Tris is undesirable, including RNA handling and nuclease inhibition by optimal pH.

Electrophysiology Buffers: Used in extracellular and intracellular solutions for patch clamp and ion channel studies at near-physiological pH.

Transfection and Electroporation: Maintains pH in delivery buffers to improve cell viability and transfection efficiency.

Cell Lysis and Extraction: Buffers lysis and wash solutions for preserving protein conformation and activity during purification.

Native PAGE and IEF: Employed in HEPES-based buffer systems for native electrophoresis and isoelectric focusing.

Bacterial Culture Media: Buffers defined and complex media (e.g., M9, LB) for Escherichia coli and other microbes to minimize pH drift during growth.