This chemical is widely used as a substrate in enzymatic assays to measure the activity of alkaline phosphatase enzymes. It is particularly valuable in biochemical and biomedical research for detecting and quantifying enzyme activity in various biological samples. When the enzyme acts on this substrate, it produces a yellow-colored product, 4-nitrophenol, which can be easily measured using spectrophotometry. This makes it a convenient tool for high-throughput screening and diagnostic applications, such as monitoring liver function or detecting bone disorders. Additionally, it is employed in ELISA (Enzyme-Linked Immunosorbent Assay) techniques for detecting antibodies or antigens in immunology studies. Its stability and sensitivity make it a reliable choice for laboratory diagnostics and research purposes.