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A Homogenous Transcreener® UDP² TR-FRET Glycosyltransferase Assay for High Throughput Screening

Transcreener UDP2 Glycosyltransferase Assay

The Transcreener UDP2 Glycosyltransferase Assay directly measures UDP produced by glycosyltransferase. These UDP measurements allow researchers to effectively determine the activity of the glycosyltransferase enzyme. The assay provides a powerful tool to screen compound libraries for glycosyltransferase modulators to help find new therapies for diseases. 

The kit comes complete with the detection reagents required to measure activity. Glycosyltransferase enzyme is not included in the inhibitor screening assay kit. Please contact us for questions related to acquiring the enzyme. This assay is designed to be used with purified enzyme preparations. Transcreener assays are not validated with cell lysates, blood, serum, or other biological samples.

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.

How Does This Glycosyltransferase Assay Work?

The Transcreener Glycosyltransferase Assay determines glycosyltransferase enzyme activity by directly measuring the UDP formed by the enzyme (i.e. without coupling enzymes) using the Transcreener UDP Assay. By detecting UDP output, the assay provides a universal method to assess the activity of any UDP-producing enzyme in real-time.

Transcreener Glycosyltransferase Assay technology uses a simple but highly effective method that consists of an antibody selective to UDP over UDP sugar donors and a far-red fluorescent tracer. UDP produced in the reaction competes with the tracer changing the fluorescent properties and providing fluorescent readout.

The glycosyltransferase assay is available with FP, FI, and TR-FRET. It is a simple mix-and-read format. Perform your enzyme reaction, add the detection reagent, and measure. The simplicity of the system yields robust results that also make it extremely amiable to HTS.

Direct Detection of UDP to Measure Glycosyltransferase Enzymatic Activity

Fluorescent Polarization (FP)

Transcreener UDP Glycosyltransferase Assay FP Readout

The workhorse. Used in many large screens. Best deck and signal stability.

Fluorescent Intensity (FI)

Transcreener UDP Glycosyltransferase Assay FI Readout

Positive FI signal. Compatible with simple fluorescence plate readers. Faster read time than FP or TR-FRET.

Time-Resolved Förster Resonance Energy Transfer (TR-FRET)

Transcreener UDP Glycosyltransferase Assay TR-FRET Readout

For customers who prefer TR-FRET detection. Uses the same filter set as HTRF®.

Applications

  • Measure Enzymatic Activity of Glycosyltransferases
  • Screen Compound Libraries for Glycosyltransferase Modulators
  • Quantify Inhibitor Potency
  • Inhibitor Selectivity Profiling
  • Measure Drug-Target Residence Time

Features

  • Direct detection of unlabeled UDP
  • Easy to use, homogenous, one-step format
  • Robust Assay Z'>0.7 under initial velocity conditions
  • Far-red fluorescent readouts minimize compound interference
  • A safe, non-radioactive method
  • Available in FP, FI, or TR-FRET readouts

Target Class Examples

  • Glucosyltransferase
  • Galactosyltransferase
  • N-acetylglucosyltransferase
  • N-acetylglucosaminyltransferase
  • Xylotransferases
  • Glucoslceramide synthase

UDP Donors

  • UDP-glucose
  • UDP-galactose
  • UDP-GlcNAc
  • UDP-GalNAc
  • UDP-xylose
  • UDP-glucuronic acid

Easy-to-Use, Mix-and-Read, HTS-Ready Assay

Run your enzyme reaction, add Transcreener reagents, and read your plates. The glycosyltransferase assay is compatible with 96, 384, and 1536-well formats.

Robust, HTS-Compatible Method

Use with Diverse Donor and Acceptor Substrates

Glycosyltransferase Assay Examples with Different Enzymes

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.

Outstanding Deck and Signal Stability

Deck stability demonstrates the ability of the reagents to remain functional after pre-mixing and awaiting dispensing. Plate stability shows the consistency of reagent signal to be read post assay completion. The stability of the reagents provides high quality data and simplifies automation for high throughput screens.

Glycosyltransferase Assay Services

Interested in moving your program forward, but don't want to bring an assay in-house? Our scientists can help! BellBrook scientists will use their extensive biochemistry and enzymology expertise to work with you and accelerate your glycosyltransferase program.

Assay Development Services

Lead Discovery Services Include:

  • Inhibitor Screening – To identify or confirm activity with the target.
  • Inhibitor Potency Profiling – Dose-response with target and/or related proteins. Fast IC50 results.
  • Residence Time Measurements – Determination of koff using ‘jump dilution’ enzymatic assay method.
  • Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.

Contact Us to Learn More

Please fill out the form below. We will respond quickly to get the conversation moving and learn how we can help. We keep things discrete, confidential, and professional.

Far Red FP, FI, & TR-FRET Readouts Validated on Major Multimode Readers

SupplierInstrumentFP AssaysFI AssaysTR-FRET Assays
TriStar2S LB 942In ReviewClick Me ArrowValidatedClick Me ArrowValidated
Tristar 5In ReviewClick Me ArrowValidatedIn Review
Mithras2 LB 943In ReviewClick Me ArrowValidatedClick Me ArrowValidated
CytationTM 5ValidatedValidatedValidated
CytationTM 3ValidatedValidatedValidated
CytationTM 1ValidatedValidatedValidated
SynergyTM H1ValidatedValidatedValidated
SynergyTM 2/H4/4Click Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
SynergyTM HTXNot CapableValidatedNot Capable
SynergyTM Neo 2ValidatedValidatedValidated
POLARstar® OmegaClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
FLUOstar® OmegaNot CapableClick Me ArrowValidatedClick Me ArrowValidated
PHERAstar® FSXClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
PHERAstar® Plus/FSClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
CLARIOstar® /PlusClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
VANTAstarTMClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
SenseIn ReviewClick Me ArrowValidatedIn Review
Analyst® GT/HTClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Gemini® XPS/EMNot CapableClick Me ArrowValidatedNot Capable
SpectraMax® M2/M2eNot CapableClick Me ArrowValidatedNot Capable
SpectraMax® M5/M5e/FlexStation® 3Not CapableClick Me ArrowValidatedClick Me ArrowValidated
SpectraMax® ParadigmClick Me ArrowValidatedClick Me ArrowContact UsClick Me ArrowContact Us
SpectraMax® iD3/iD5Click Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
SpectraMax® i3xClick Me ArrowValidatedIn ReviewClick Me ArrowValidated
EnVision®/EnVision® XciteClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Infinite® M1000/M1000Pro/Safire2TMClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Infinite® M200Not CapableClick Me ArrowValidatedNot Capable
Infinite® F500Click Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Infinite® F200/UltraevolutionClick Me ArrowContact UsClick Me ArrowValidatedClick Me ArrowContact Us
SparkTM 10MClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated

What's Included

What You Will Need

Transcreener UDP² Glycosyltransferase Assay Kits

* For custom or bulk orders (over 100,000 wells) please contact us (info@bellbrooklabs.com) for a quote.

Assay Plates for FI & FP

Assay Plates for TR-FRET

Glycosyltransferases As Drug Targets

Glycosyltransferases (GTs) are catalysts for the transfer of a nucleotide sugar donor to a glycosyl acceptor substrate. Glycosyltransferases play roles in a variety of biochemical processes, including cellular adhesion, cell signaling, and bacterial biosynthesis. Because of their essential parts in these processes, glycosyltransferases have gained large interest for drug discovery (Wagner 2010).

Glycosyltransferase inhibitors are often targeted for metabolic disorders. Continuing to build an understanding of GT enzymes is step one. Finding therapeutic methods to reign in overactivity requires selective technologies that can help assess the ability to interact with the target. Robust, HTS-ready assays that are accurate yet flexible provide a powerful tool to accelerate discovery.

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