N-Succinyl-Ala-Ala-Pro-Phe-7-amido-4-methylcoumarin is primarily used as a fluorogenic substrate in enzymatic assays, particularly for detecting and measuring the activity of proteases such as chymotrypsin and subtilisin. Its application is crucial in biochemical research, where it helps in understanding enzyme kinetics and mechanisms. Upon enzymatic cleavage, it releases a fluorescent product, 7-amino-4-methylcoumarin, which can be quantitatively measured using fluorescence spectroscopy. This property makes it a valuable tool in high-throughput screening for identifying protease inhibitors, which are potential candidates for therapeutic drugs. Additionally, it is employed in cell biology studies to investigate protease activity within cellular processes and pathways. Its sensitivity and specificity make it a preferred choice for both in vitro and in vivo studies related to protease function and regulation.