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GlmU (UDP-N-acetylglucosamine pyrophosphorylase) is a bifunctional enzyme. It adds acetyl and uridyl groups to glucosamine-1-phosphate to form UDP-GlcNAc, an essential peptidoglycan precursor. UDP-GlcNAc is also used to build the N-acetylglucosamine polysaccharide adhesin that many bacteria use to form biofilms.
Pack Sizes
Both sizes include E. coli GlmU and pyrophosphatase. The only difference is the number of assays: GLU100KE provides reagents for 100 assays and GLU500KE for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (GLU100KE) | 100 assays; enzyme included |
|---|---|
| 500 assays (GLU500KE) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (GLU100KE) | 500 assays (GLU500KE) |
|---|---|---|
| 10 x Buffer | 400 µl | 2 ml |
| 100 x Enzyme Substrate | 35 µl | 170 µl |
| 100 x E. coli GlmU (500 nM) | 35 µl | 170 µl |
| 100 x Pyrophosphatase (PPase, 10 U/ml) | 35 µl | 170 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 5 ml | 25 ml |
The 100 x Enzyme Substrate contains 2.5 mM glucosamine-1-phosphate, 2.5 mM acetyl CoA and 2.5 mM UTP.
Assay Principle
GlmU releases pyrophosphate, which the included pyrophosphatase converts to inorganic phosphate. The phosphate is detected with a dye by absorbance at 650 nm, so the signal tracks GlmU activity and drops with inhibitors.
Specifications
| Assay Target | E. coli GlmU activity |
|---|---|
| Target Type | Pyrophosphorylase |
| Organism | E. coli |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Colorimetric |
| Detection Wavelength | Absorbance 650 nm |
| Plate Format | 384/96-well plate or cuvette |
| Instrument Required | Plate reader or spectrophotometer |
| Assay Time | About 65 min (60 min reaction + 5 min detection) |
| Linearity Test | Start reactions at different time points and plot signal vs. time to define the linear range |
| Sizes | 100 assays, 500 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- E. coli GlmU Assay (PDF, see Documents): Covers catalog # GLU100KE, GLU500KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli GlmU activity assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.