This compound is primarily used in biochemical research as a substrate for proteolytic enzymes, particularly serine proteases like thrombin and trypsin. It helps in studying enzyme kinetics and activity by releasing p-nitroaniline upon cleavage, which can be measured spectrophotometrically. This makes it valuable in assays for understanding enzyme mechanisms and screening protease inhibitors. It is also utilized in diagnostic applications to detect and quantify protease activity in biological samples. Additionally, it aids in the development of therapeutic agents targeting protease-related diseases.