Bio-Techne Corporation
Methods and kits for assaying endoglycosidase activity

Abstract:

A method for assaying endoglycosidase activity includes providing a proteoglycan having a glycosaminoglycan chain with a non-reducing end; treating the proteoglycan with a glycosyltransferase to incorporate a carbohydrate into the non-reducing end of the glycosaminoglycan chain, wherein the carbohydrate includes a click chemistry moiety; adding a label to the proteoglycan, wherein the label includes a click chemistry moiety that reacts to the click chemistry moiety of the carbohydrate such that the label attaches to the carbohydrate to form a labeled proteoglycan; immobilizing the labeled proteoglycan on a multi-well plate, wherein the multi-well plate includes a specific anti-proteoglycan antibody for binding the labeled proteoglycan; treating the labeled proteoglycan with an endoglycosidase specific to the glycosaminoglycan chain; and detecting the labeled proteoglycan.