Protein Electrophoresis (Bis-Tris Gels): Serves as the buffering component for SDS-PAGE and native PAGE systems (with MES or MOPS running buffers) to ensure sharp, pH-stable protein separation.

Western Blotting: Maintains stable pH during gel-to-membrane protein transfer and antibody incubation when using Bis-Tris gel systems.

Protein Purification: Used in ion-exchange and affinity chromatography to maintain a stable pH 6.5 environment during binding and elution.

Enzyme Assays: Provides reliable buffering at pH 6.5 for enzymatic reactions and kinetic measurements.

Isoelectric Focusing (IEF): Buffers proteins in the mildly acidic range to support precise focusing based on isoelectric points.

Protein Storage and Stabilization: Used for formulation, dialysis, and storage to maintain protein integrity at pH 6.5.

Sample Preparation and Buffer Exchange: Employed for dilution, desalting, and dialysis where a defined pH 6.5 buffer is required.