Enzyme Activity of Glycolipid Specific Glycosyltransferase Enzymes GM2/GD2 Synthase β1,4-GalNAc Transferase (B4GALNT1).

Measurement of enzyme activity of glycosyltransferases enables the direct observation of the reaction of substrate specificity, enzyme kinetics, and interaction with other molecules. Cell-membrane fractionations containing glycosyltransferases are collected by nitrogen cavitation, and are utilized for enzyme reaction in cell-free glass tubes. Thin-layer chromatography is used to separate reaction products, and the products are visualized by an imaging analyzer. Here, we describe the whole method in detail and step by step.