UMP/CMP-Glo™ Glycosyltransferase Assay

Universal Glycosyltransferase Activity Assay

  • Easy, "add-incubate-read" protocol
  • Detects sialyltransferases and phosphoglycosyltransferases that use CMP-, CDP- or UDP-sugars as donor substrates
  • Ideal for low-activity glycosyltransferases

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Catalog number selected: VA1130

$ 385.00
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UMP/CMP-Glo™ Glycosyltransferase Assay
200 assays
$ 385.00
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A Simple, Bioluminescent Assay for Glycosyltransferase Activity

The UMP/CMP-Glo™ Glycosyltransferase Assay is a homogeneous, single-reagent-addition method to rapidly detect UMP or CMP formation in glycosyltransferase reactions. After the glycosyltransferase reaction, an equal volume of UMP/CMP Detection Reagent is added to simultaneously convert the UMP or CMP 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 UMP or CMP from low nM to 50μM. Luminescence can be correlated to UMP or CMP concentration by using a UMP or CMP standard curve.

Add-Mix-Read for Results

Start with purified enzymes, sugar donors and acceptor molecules, and you can detect the reaction products UMP or CMP from phosphoglycosyltransferases or sialyltransferases, respectively. Just add the UMP/CMP Detection Reagent and measure luminescence.

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Works with Many Sialyltransferases and Phosphoglycosyltransferases

The UMP/CMP-Glo™ Assay can be used to measure the activity of just about any sialyltransferase that uses CMP-sugars or any phosphoglycosyltransferase that uses UDP-sugars as a substrate. Here, we looked at sialyltransferases involved in protein O-glycosylation.

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Ideal for Low-Activity Glycosyltransferases and Phosphoglycosyltransferases

The sensitivity of the UMP/CMP-Glo™ Glycosyltransferase Assay means it can detect low UMP or CMP 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 UMP or CMP, you can obtain a high signal-to-background ratio using the assay.

The data below show the linearity and sensitivity of the UMP/CMP-Glo™ Glycosyltransferase Assay. Here, a UMP or CMP standard curve was prepared over the indicated concentration range in 25µl of 1X glycosyltransferase reaction buffer in a solid white 96-well plate. UMP/CMP-Glo™ Glycosyltransferase Assay was performed using 25µl of UMP/CMP Detection Reagent at room temperature. Luminescence was recorded using a GloMax® luminometer. Values represent the mean of two replicates. The tables show luminescence measured 1 hour after adding the UMP/ CMP Detection Reagent, and signal-to-background ratios calculated for each concentration of the standard curve.

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Luminescent Signal is Stable Over Several Hours

The UMP/CMP-Glo™ Assay  generates a highly stable luminescent signal that is stable for more than 3 hours, making it convenient to batch-process assay plates.

The data here show that the signal obtained with the UMP/CMP-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.

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Specifications

You are viewing: VA1130 Change Configuration

What's in the box?

Item Part # Size

ATP Detection Substrate

V363C 1 × 1 vial

Nucleotide Detection Buffer

V696A 1 × 5ml

UMP, 10mM

VA113A 1 × 100μl

CMP, 10mM

VA114A 1 × 100μl

UMP/CMP-Glo™ Enzyme

VA115A 1 × 200μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

See Protocol for detailed storage recommendations.

Patents and Disclaimers

U.S. Pat. Nos. 7,741,067 and 8,361,739.

Specifications

You are viewing: VA1131 Change Configuration

What's in the box?

Item Part # Size

ATP Detection Substrate

V363A 1 × 1 vial

Nucleotide Detection Buffer

V696B 1 × 10ml

UMP, 10mM

VA113A 1 × 100μl

CMP, 10mM

VA114A 1 × 100μl

UMP/CMP-Glo™ Enzyme

VA115B 1 × 400μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

See Protocol for detailed storage recommendations.

Patents and Disclaimers

U.S. Pat. Nos. 7,741,067 and 8,361,739.

Specifications

You are viewing: VA1132 Change Configuration

What's in the box?

Item Part # Size

ATP Detection Substrate

V363B 1 × 1 vial

Nucleotide Detection Buffer

V696C 1 × 100ml

UMP, 10mM

VA113A 1 × 100μl

CMP, 10mM

VA114A 1 × 100μl

UMP/CMP-Glo™ Enzyme

VA115C 4 × 1ml

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

See Protocol for detailed storage recommendations.

Patents and Disclaimers

U.S. Pat. Nos. 7,741,067 and 8,361,739.

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