HEPES is widely used as a buffering agent in biological and biochemical research due to its ability to maintain a stable pH in cell culture media and enzymatic reactions. It is particularly favored in cell and tissue culture applications because it is non-toxic to cells and does not interfere with biochemical processes. HEPES is also commonly employed in electrophoresis, protein purification, and molecular biology techniques, where precise pH control is critical. Additionally, it is utilized in the preparation of reagents for diagnostic assays and in the study of cellular physiology, as it effectively buffers solutions in the physiological pH range (7.2–7.4). Its stability and minimal metal ion chelation make it suitable for use in experiments involving sensitive biological materials.