Enzymatic method for detecting a labelled segment and a solution or composition therefor

Abstract

A non-radioactive method of detecting the enzymes beta galactosidase or beta glucosidase directly or for the detection of a ligand and antiligand complex is provided wherein beta galactosidase or the complex labelled with beta galactosidase or a tracer having beta galactosidase conjugated thereto is reacted with 5-bromo-4-chloro-3-indolyl-B-D-galactoside and a tetrazolium salt to produce a colored formazan or a color change indicative of the presence of beta galactosidase, or wherein beta glucosidase or the complex labelled with the beta glucosidase or a tracer having beta glucosidase conjugated thereto is reacted with 5-bromo-4-chloro-3-indolyl-B-D-glucoside and a tetrazolium salt to produce a colored formazan or a color change indicative of the presence of beta glucosidase. Optionally, the galactosidase-galactoside determination or the glucosidase-glucoside determination may further include catalyst phenazine methosulfate (PMS) as a reactant. The preferred tetrazolium salt is dimethylthiazol diphenyl tetrazolium (MTT). A solution or composition of 5-bromo-4-chloro-3-indolyl-B-D-galactoside or 5-bromo-4-chloro-3-indolyl-B-D-glucoside, and a tetrazolium salt, as well as a test kit including the same, is also provided for carrying out the chromogenic method of detection.

Dependencies are inferred from the claim text.