Used as a synthetic substrate in enzymatic assays, particularly for studying serine proteases such as trypsin and thrombin. The p-nitroanilide group allows for colorimetric detection upon cleavage, enabling real-time monitoring of enzyme activity. Commonly employed in kinetic studies and inhibitor screening due to its high specificity and measurable absorbance change. Also utilized in quality control of proteolytic enzymes and in the development of diagnostic assays.