N-Acetylglucosaminyltransferase (GnT) Assays Using Fluorescent Oligosaccharide Acceptor Substrates:

Determining glycosyltransferase activities gives a clue for better understanding an underlying mechanism for glycomic alterations of carrier molecules. N -glycan branch formation is concertedly regulated by cooperative and competitive activities of N -acetylglucosaminyltransferases (GnTs). Here, we describe methods for large scale preparation of the oligosaccharide acceptor substrate, fluorescence-labeling of oligosaccharides by pyridylamination, quality control, and reversed phase HPLC-based measurement of GnT activities including GnT-III, IV, V, and IX.