Robiul Hasan Bhuiyan, Farhana Yesmin, Keiko Furukawa, Yuhsuke Ohmi, Koichi Furukawa
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.