Homogenous Transcreener® UDP² TR-FRET Assay for High Throughput Screening with Glycosyltransferases
About the Transcreener UDP2 Glycosyltransferase Assay Kits
The Transcreener UDP2 Glycosyltransferase Assay is a single step, homogenous, fluorescent assay for detection and screening of UDP-producing enzymes. Direct detection of UDP with a fluorescence polarization (FP) or TR-FRET readout provides a safe, HTS-compatible alternative to cumbersome radioassay methods and is more sensitive and less subject to interference than coupled assay methods, where the UDP is converted to another product. Based on BellBrook Lab’s proprietary nucleotide immunodetection technology, the Transcreener UDP2 Glycosyltransferase Assay is compatible with any enzyme class that produces UDP, including UDP-glucose-, UDP-galactose- and UDP-glucuronosyl-transferases as well as glycogen, hyaluranon and cellulose synthases.
Learn how researchers at the Genesis Biotechnology group used the Transcreener UDP assay in a high throughput screen to discover Clostridium difficile (C. difficile) toxin B (TcdB) inhibitors.
Direct Detection of UDP for Glycosyltransferases
Target Class Examples
- Glucosyltransferase
- Galactosyltransferase
- N-acetylglucosyltransferase
- N-acetylglucosamyltransferase
- Xylotransferases
- Glucoslceramide synthase
Applications
- Screen compound libraries for inhibitors
- Hit confirmation
- IC50 Determination
- Determine inhibitor selectivity between enzymes
- Measure glycosyltransferase activity
Simple, “Mix-and-Read” Assay Format
Use with Diverse Donor and Acceptor Substrates
GALNT2, GALNT3, BGalT1, and Toxin B substrates were titrated in 4 buffers. Raw data converted to product formation show a linear correlation of the enzyme with the UDP product, confirming initial velocity conditions.
UDP Donors
- UDP-glucose
- UDP-galactose
- UDP-GlcNAc
- UDP-GalNAc
- UDP-xylose
- UDP-glucuronic acid
Robust, HTS-Compatible Assay
Outstanding Deck and Signal Stability
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One Way to Accelerate Biochemical Assay Development
Download Case Study
Discovery of GALNT3 Inhibitors Using a Universal Glycosyltransferase Assay and Orthogonal Pooled Screening
