H-Ile-AMC (Isoleucine-7-amido-4-methylcoumarin) is a fluorogenic peptide substrate consisting of the amino acid isoleucine conjugated to the fluorophore 7-amino-4-methylcoumarin (AMC). This compound is widely used as a specific and sensitive substrate for various proteolytic enzymes, particularly serine proteases. Upon enzymatic cleavage of the amide bond between the isoleucine residue and the AMC moiety, the highly fluorescent AMC is released, resulting in a significant increase in fluorescence intensity. Its primary application is in biochemical and biomedical research for the continuous, real-time assay of protease activity. It is extensively utilized to study enzymes such as dipeptidyl peptidase IV (DPP-IV), prolyl oligopeptidase, and other aminopeptidases. The assay format is crucial for high-throughput screening of enzyme inhibitors, kinetic studies, and characterizing protease function in various biological pathways. Due to its specificity and the sensitive detection method, H-Ile-AMC is a key reagent in the development of diagnostic assays and in drug discovery programs targeting proteases involved in diseases like diabetes, cancer, and neurological disorders. It is supplied as a stable solid and typically dissolved in DMSO or DMF for use in assay buffers.