Chemiluminescent acridan phosphate labelling compounds for detection in gels

We have developed a chemiluminescent acridan phosphate labelling compound which produces a flash of light upon chemical triggering. Sequential treatment of the label with acid and a strong base, one of which contains a peroxide, initiates light emission as a rapid flash lasting 1--2 s. Labelling of analytes with these compounds permits their direct detection in a non-enzymatic assay format. Representative compounds have been linked to BSA as a model protein analyte and detected by chemiluminescence assay. Light intensity correlated with the amount of BSA over six orders of magnitude, permitting the detection of 50 amol of protein in solution. We have also demonstrated for the first time that labelled proteins can be separated by electrophoresis without destruction of the label and that chemiluminescence can be produced and detected directly in the gel. The ease, speed and sensitivity of detection by this new method should enable the development of simpler assays for a wide variety of analytes.