GALNT3 Assay – Transcreener Application

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GALNT3 Assay - A Transcreener UDP2 Assay Application

Transcreener UDP² Assay directly measures UDP produced by GALNT3 (Polypeptide N-Acetylgalactosaminyltransferase 3). These UDP measurements allow researchers to effectively determine the activity of the GALNT3 enzyme. The assay provides a powerful tool to screen compound libraries for GALNT3 modulators to help find new therapies for disease. 

The kit comes complete with the detection reagents required to measure activity. GALNT3 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.

How Does This GALNT3 Assay Work?

The Transcreener GALNT3 Assay determines GALNT3 enzyme activity by directly measuring the UDP formed by the enzyme 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 GALNT3 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 GALNT3 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 GALNT3 Enzymatic Activity

Fluorescent Polarization (FP)

Transcreener UDP GALNT3 Assay Schematic

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 GALNT3
  • Screen Compound Libraries for GALNT3 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

Easy-to-Use, Mix-&-Read, HTS-Ready

Run your enzyme reaction, add Transcreener reagents, and read your plates. The GALNT3 assay is compatible with 96, 384, and 1536-well formats.

Transcreener Mix and Read Assay

Robust Assay Yields Quality Data

Z’ measurements using optimized GALNT3 reaction conditions indicate a robust assay. Robust data is vital for sizeable high throughput screens that are difficult to complete due to massive sample quantity.

GALNT3 Assay Robustness Z'
Z' GALNT3 Data Values

UDP-GalNAc/UDP Standard Curve

The standard curve mimics an enzyme reaction (as UDP-sugar concentration decreases, UDP concentration increases). Z’ at 10% conversion (>0.85) shows excellent assay sensitivity under initial velocity conditions. The lower limit of detection is 100 nM UDP. Data shown here and for the following assays use the FI format. The UDP² Antibody-IRDye QC-1  concentration in the 10 µL reaction mixture is 1.049 µg/mL.

10 μM UDP/UDP-GalNAc Standard Curve

UDP-GalNAc Standard Curve
UDP-GalNAc Standard Curve Values

Detection of UDP Under GALNT3 Initial Velocity Conditions

GALNT3 was titrated in the presence and absence of Mucin 10-EA2 peptide (10 μM) and UDP-GalNAc (10 μM) for 60 minutes at room temperature. Data is converted to UDP formed using the UDP-GalNAc/UDP standard curve. The assay demonstrates a linear correlation under initial velocity conditions when raw data is converted to UDP using a standard curve. 

GALNT3 Enzyme Titration

GALNT3 Enzyme Titration

Linear Response

GALNT3 Assay Linear Response

Time Course - Detection of UDP Released by GALNT3

Transcreener detection reagents were added to enzyme reaction prior to starting reactions by addition of UDP-GalNAc. Plates were then read at 30 minute intervals. Raw data in relative fluorescence units. Assay shows a linear correlation when converted to UDP formed.

GALNT3 Time Course Titration
GALNT3 Assay Time Course Standard Curve

GALNT3 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 GALNT3 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 ReviewIn Review
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 Assay Kits - For GALNT3 Activity Assays

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

Cat. No.Product DescriptionPriceAdd to Cart
3019-ATranscreener UDP2 FI Assay (200 Assay, 96 Well)$359.00Add to Cart
3019-1KTranscreener UDP2 FI Assay (1,000 Assay, 384 Well)$759.00Add to Cart
3019-10KTranscreener UDP2 FI Assay (10,000 Assay, 384 Well)$4,299.00Add to Cart
3019-100KTranscreener UDP2 FI Assay (10 x 10,000 Assay, 384 Well)$21,495.00Add to Cart
3018-1KTranscreener UDP2 FP Assay (1,000 Assay, 384 Well)$725.00Add to Cart
3018-10KTranscreener UDP2 FP Assay (10,000 Assay, 384 Well)$4,099.00Add to Cart
3018-100KTranscreener UDP2 FP Assay (10 X 10,000 Assay, 384 Well)$20,495.00Add to Cart
3022-1KTranscreener UDP2 TR-FRET Assay (1,000 Assay, 384 Well)$789.00Add to Cart
3022-10KTranscreener UDP2 TR-FRET Assay (10,000 Assay, 384 Well)$4,449.00Add to Cart
3022-100KTranscreener UDP2 TR-FRET Assay (10 X 10,000 Assay, 384 Well)$22,245.00Add to Cart

Assay Plates for FI & FP

Cat. No.Product DescriptionPriceAdd to Cart
2233Corning 96-Well Black Assay Plates (#3686), 3-Pack (200+ Assays)$55.00Add to Cart
2229Corning 384-Well Black Assay Plates (#4514), 3-Pack (1,000+ Assays)$60.00Add to Cart
2230Corning 384-Well Black Assay Plates (#4514), 30-Pack (10,000+ Assays)$479.00Add to Cart

Assay Plates for TR-FRET

Cat. No.Product DescriptionPriceAdd to Cart
2234Corning 96-Well White Assay Plates (#3642), 3-Pack (200+ Assays)$55.00Add to Cart
2231Corning 384-Well White Assay Plates (#4513), 3-Pack (1,000+ Assays)$65.00Add to Cart
2232Corning 384-Well White Assay Plates (#4513), 30-Pack (10,000+ Assays)$529.00Add to Cart

The Role of GALNT3 As a Therapeutic Target

Glycosyltransferases catalyze the transfer of sugar molecules to protein, lipid, and carbohydrate acceptors along with endogenous and xenobiotic small molecules. More than 200 human glycosyltransferases are known to exist and have diverse metabolic and regulatory roles. Of those, at least 20 distinct GALNTs (Polypeptide N-Acetylgalactosaminyltransferases) are found in the Golgi.

Since GALNTs catalyze the initial step of mucin-type O-glycosylation by transferring GalNac to Thr or Ser residues, these enzymes are targets for cancer interrogation. Multiple cancers directly link to GALNT overexpression and dysregulation. The Transcreener UDP GALNT3 Assay is an excellent tool for researchers examining the therapeutic effects of GALNT3.

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1232 Fourier Drive, Suite 115
Madison, Wisconsin 53717 USA
(608) 443-2400

info@bellbrooklabs.com

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