2-Iminobiotin hydrobromide is primarily used in biochemical research as a competitive inhibitor of biotin-dependent enzymes. It is particularly valuable in studies involving avidin-biotin systems, where it helps to investigate binding interactions and enzyme mechanisms. This compound is also employed in affinity chromatography to purify biotin-binding proteins, enabling the isolation and analysis of specific biomolecules. Additionally, it serves as a tool in molecular biology to study the role of biotin in cellular processes and metabolic pathways. Its application extends to diagnostic assays, where it aids in the detection and quantification of biotin-related interactions.