Abstract
The use of 1, 2-dioxetane chemiluminescent enzyme substrates, including AMPPD®, CSPD®, CDP® and CDP-Star® for alkaline phosphatase and Galacton-Star® substrate for β-galactosidase, provides highly sensitive detection for numerous immunoassay and nucleic acid detection formats. Enzyme cleavage of the 1, 2-dioxetane substrate generates a metastable anion intermediate that decomposes with the concomitant emission of light. Light emission exhibits glow kinetics, enabling the use of multiple imaging platforms for signal detection, including film-based, luminometers, low-light sensitive camera and phosphor screen instrumentation systems. Applications include both membrane-based immunodetection of proteins and nucleic acid blot hybridization, and solution-based immunoassays and nucleic acid capture/hybridization assays performed in a microwell plate.
