Glycosyltransferase Activity Kit

View Full Assay Principle

The Glycosyltransferase Activity Kit (Catalog # EA001) provides a simple, non-radioactive high-throughput compatible format for assaying the enzyme activity of all glycosyltransferases that use diphosphonucleotide sugars as donor substrates. This kit utilizes the nucleotidase ENTPD3/CD39L3 as a coupling phosphatase to remove inorganic phosphate quantitatively from the leaving nucleotide diphosphate, such as UDP or GDP, generated during glycosyltransferase reactions.

Features

• Applicable for all glycosyltransferases that generate leaving groups with a removable phosphate
• Non-radioactive
• No time-consuming separation steps (e.g. column chromatography)
• Provides accurate kinetic analysis of glycosyltransferase reactions
• Amenable to high-throughput analysis

Kit Contents

• NS0-expressed Recombinant Human ENTPD3/CD39L3
• UDP
• Phosphatase Buffer 1
• 100 mM MnCl₂
• Phosphate Standard
• Malachite Green Reagents A & B

Data Example

Activity of B4GalT1

 

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The ability of Recombinant Human beta-1,4-Galactosyltransferase 1/B4GalT1 (Catalog # 3609-GT) to transfer galactose from UDP-galactose to N-acetyl-alpha-D-glucosamine was measured using the Glycosyltransferase Activity Kit (Catalog # EA001). The activity was plotted against enzyme input, and the specific activity of B4GalT1 was determined to be 1806 pmol/min/µg.