Homogeneous Detection of Glycosyltransferase Activities with Universal Bioluminescent Assays

Part # PS295

Abstract

Hicham Zegzouti, Laurie Engel, Jacquelyn Hennek, Juliano Alves, Gediminas Vidugiris, and Said Goueli
Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711

We developed four bioluminescent assays for measuring GT activities based on UDP, GDP, UMP and CMP quantification. These assays have the following features:
• One-step detection: After GT reaction, the nucleotide product simultaneously converted to ATP, then to light in a robust luciferase reaction.
• The light output is proportional to the nucleotide concentration ranging from low nM to 25-50μM.
• Very sensitive and robust assays, two highly desirable and essential features required for measuring the activity of the majority of GT classes.
• Simple assays, do not require antibodies, nor modified substrates.

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