Used primarily in bioluminescent assays and detection systems due to its ability to emit light upon enzymatic activation. The compound serves as a chemiluminescent substrate in immunoassays and nucleic acid detection, where alkaline phosphatase enzyme removes the phosphate group, triggering decomposition of the dioxetane ring and producing light. This light emission allows for highly sensitive, real-time detection of biological targets without the need for radioactive labels. Commonly found in diagnostic kits, including ELISA and blotting applications, it enables low-background, high-signal detection ideal for high-throughput screening and clinical diagnostics.