UDP-Glo™ Glycosyltransferase Assay Technical Manual
Literature # TM413
The UDP-Glo™ Glycosyltransferase Assay is a bioluminescent assay for detecting the activity of glycosyltransferases that use UDP-sugars as donor substrates and release UDP as a product. Glycosyltransferases transfer sugar from a nucleotide-glycosyl donor (e.g., UDP-Galactose, UDP-Glucose, UDP-GlcNAc, UDP-GalNAc and UDP-Glucuronic acid) to an acceptor molecule. Other sugars are typical glycosyltransferase acceptor substrates but can also be lipids, proteins, nucleic acids, xenobiotics or other small molecules. In a glycosyltransferase reaction, the UDP moiety is released as a product; therefore, an assay that detects UDP would be suitable for monitoring the activity of the majority of glycosyltransferases.
The UDP-Glo™ Glycosyltransferase Assay is a homogenous, single-reagent-addition method to rapidly detect UDP formation in glycosyltransferase reactions. After the glycosyltransferase reaction, an equal volume of UDP Detection Reagent is added to simultaneously convert the UDP product to ATP and generate light in a luciferase reaction. The light output is proportional to the concentration of UDP from low nM to 25µM. The assay is fast, simple and highly sensitive, features that are desirable and essential for measuring the activity of different UDP-sugar-utilizing glycosyltransferases covering the majority of glycosyltransferase classes. Therefore, the UDP detection assay uses less enzyme in glycosyltransferase reactions. The UDP-Glo™ Assay is performed in a single well of a multiwell plate, and can be used to detect glycosyltransferase activity in as little as a 5µl reaction.
Summary of Changes
Added two new components, UDP-Glo™ Enzyme and Enzyme Dilution Buffer, to replace UDP-Glo™ Solution, and updated images and instructions about making the UDP-Glo™ working solution.
Printed in USA. Revised 11/15.