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| 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. |