4-Methylumbelliferyl phosphate is widely used as a substrate in enzymatic assays, particularly for detecting the activity of alkaline phosphatase. When the enzyme acts on this compound, it releases 4-methylumbelliferone, a highly fluorescent product, allowing for sensitive and quantitative measurements. This application is crucial in biomedical research, clinical diagnostics, and biochemistry for studying enzyme kinetics, monitoring enzymatic reactions, and detecting biomarkers in biological samples. Additionally, it is employed in histochemistry and immunohistochemistry to visualize and localize enzyme activity in tissues. Its high sensitivity makes it a valuable tool in high-throughput screening and diagnostic assays.