GDP-Glo™ Glycosyltransferase Assay
Universal Glycosyltransferase Activity Assay
- Easy, "add-incubate-read" protocol
- Detects any glycosyltransferase that uses GDP-sugar as a substrate
- Ideal for low-activity glycosyltransferases
Catalog Number:
Size
Add a GDP-sugar substrate
Catalog Number: VA1090
Catalog Number: VA1091
Catalog Number: VA1092
Catalog Number: VA1093
Catalog Number: VA1094
Catalog Number: VA1095
Catalog Number: VA1096
A Simple, Bioluminescent Assay for Glycosyltransferase Activity
The GDP-Glo™ Glycosyltransferase Assay is a homogeneous, single-reagent-addition method to rapidly detect GDP formation in glycosyltransferase reactions. After the glycosyltransferase reaction, an equal volume of GDP Detection Reagent is added to simultaneously convert the GDP product to ATP and generate light in a luciferase reaction. The light generated is detected using a luminometer, and the light output is proportional to the concentration of GDP from low nM to 25μM. Luminescence can be correlated to GDP concentration by using a GDP standard curve.
Add-Mix-Read for Results
Start with purified enzymes, sugar donors and acceptor molecules, and you can detect the glycosyltransferase reaction product GDP. Just add the GDP Detection Reagent and measure luminescence.
Works with Most Glycosyltransferases
The GDP-Glo™ Assay can be used to measure the activity of just about any glycosyltransferase that uses GDP-sugar as a substrate. Here, we looked at Fucosyltransferases involved in antigen display on immune cells.
Detects Low Nanomolar Amounts of GDP; Ideal for Low-Activity Glycosyltransferases
The sensitivity of the GDP-Glo™ Glycosyltransferase Assay means it can detect low GDP concentrations with high dynamic range. Therefore, whether assaying for a low-activity glycosyltransferase whose sugar transfer rate is low or using a low concentration of enzyme that produces a small amount of GDP, you can obtain a high signal-to-background ratio using the assay.
The data below show the linearity and sensitivity of the GDP-Glo™ Glycosyltransferase Assay. Here, a GDP standard curve was prepared over the indicated concentration range in 25µl of 1X glycosyltransferase reaction buffer in a solid white 96-well plate. GDP-Glo™ Glycosyltransferase Assay was performed using 25µl of GDP Detection Reagent at room temperature. Luminescence was recorded using a GloMax® luminometer. Values represent the mean of four replicates. The table shows luminescence measured 1 hour after adding the GDP Detection Reagent, and signal-to-background ratio calculated for each concentration of the standard curve.
Luminescent Signal is Stable Over Several Hours
The GDP-Glo™ Assay generates a highly stable luminescent signal that is stable for more than 3 hours, making it convenient to batch-process multiple assay plates.
The data here show that the signal obtained with the GDP-Glo™ Assay is stable for several hours. To determine signal stability, luminescence was recorded at 1 and 3 hours after adding detection reagent. The signal-to-background ratio did not change over the time measured (data not shown). RLU = relative light units.
Protocols
Specifications
Catalog Number:
What's in the box
| Item | Part # | Size |
|---|---|---|
ATP Detection Substrate |
V363C | 1 × 1 vial |
Nucleotide Detection Buffer |
V696A | 1 × 5ml |
Enzyme Dilution Buffer |
V773A | 1 × 1.5ml |
GDP, 10mM |
VA111A | 1 × 100μl |
GDP-Glo™ Enzyme |
VA112A | 1 × 20μl |
SDS
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Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Size |
|---|---|---|
|
ATP Detection Substrate |
V363A | 1 × 1 vial |
Nucleotide Detection Buffer |
V696B | 1 × 10ml |
Enzyme Dilution Buffer |
V773A | 2 × 1.5ml |
GDP, 10mM |
VA111A | 1 × 100μl |
GDP-Glo™ Enzyme |
VA112B | 1 × 40μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Size |
|---|---|---|
|
ATP Detection Substrate |
V363B | 1 × 1 vial |
Nucleotide Detection Buffer |
V696C | 1 × 100ml |
Enzyme Dilution Buffer |
V773B | 1 × 15ml |
GDP, 10mM |
VA111A | 1 × 100μl |
GDP-Glo™ Enzyme |
VA112C | 1 × 200μl |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
ATP Detection Substrate |
V363C | 1 × 1 vial | ||
Nucleotide Detection Buffer |
V696A | 1 × 5ml | ||
Enzyme Dilution Buffer |
V773A | 1 × 1.5ml | ||
GDP, 10mM |
VA111A | 1 × 100μl | ||
GDP-Glo™ Enzyme |
VA112A | 1 × 20μl | ||
GDP-Fucose |
VA109A | 1 × 50μl | 50mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
|
ATP Detection Substrate |
V363A | 1 × 1 vial | ||
Nucleotide Detection Buffer |
V696B | 1 × 10ml | ||
Enzyme Dilution Buffer |
V773A | 2 × 1.5ml | ||
GDP, 10mM |
VA111A | 1 × 100μl | ||
GDP-Glo™ Enzyme |
VA112B | 1 × 40μl | ||
GDP-Fucose |
VA109A | 1 × 50μl | 50mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
ATP Detection Substrate |
V363C | 1 × 1 vial | ||
Nucleotide Detection Buffer |
V696A | 1 × 5ml | ||
Enzyme Dilution Buffer |
V773A | 1 × 1.5ml | ||
GDP, 10mM |
VA111A | 1 × 100μl | ||
GDP-Glo™ Enzyme |
VA112A | 1 × 20μl | ||
GDP-Mannose |
VA110A | 1 × 50μl | 100mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
|
ATP Detection Substrate |
V363A | 1 × 1 vial | ||
Nucleotide Detection Buffer |
V696B | 1 × 10ml | ||
Enzyme Dilution Buffer |
V773A | 2 × 1.5ml | ||
GDP, 10mM |
VA111A | 1 × 100μl | ||
GDP-Glo™ Enzyme |
VA112B | 1 × 40μl | ||
GDP-Mannose |
VA110A | 1 × 50μl | 100mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
Resources
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Frequently Used With
Ultra Pure GDP-Sugar Substrates
Substrates for use with the GDP-Glo™ Glycosyltransferase Assay.
VA1097, VA1098, VA1099, VA1100
GloMax® Explorer System
Easy-to-use, flexible multimode detection system.
GM3500, GM3510
GloMax® Discover System
High-performance microplate reader for detecting luminescence, fluorescence and absorbance.
GM3000
GloMax® Navigator System
Dedicated microplate luminometer.
GM2000, GM2010
