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Bacterial peptidoglycan biosynthesis uses four related amino acid-adding enzymes, and MurC (UDP-N-acetylmuramic acid:L-alanine ligase) acts first. It joins L-alanine to UDP-MurNAc to give UDP-MurNAc-L-Ala, while ATP is hydrolyzed to ADP and inorganic phosphate. Because the enzyme is essential, it is an attractive antibacterial drug target.
Pack Sizes
Both sizes include the E. coli MurC enzyme. The only difference is the number of assays: MURC100KE is for 100 assays and MURC500KE for 500 assays (384-well format), with component volumes scaled.
| 100 assays (MURC100KE) | 100 assays; enzyme included |
|---|---|
| 500 assays (MURC500KE) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (MURC100KE) | 500 assays (MURC500KE) |
|---|---|---|
| 10 x Assay buffer | 400 µl | 2 ml |
| 100 x UDP-MurNAc | 35 µl | 170 µl |
| 100 x L-Ala | 35 µl | 170 µl |
| 100 x ATP | 35 µl | 170 µl |
| 100 x E. coli MurC (2000 nM) | 35 µl | 170 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 5 ml | 25 ml |
Assay Principle
The kit measures the inorganic phosphate released when MurC adds L-alanine to UDP-MurNAc and hydrolyzes ATP with a phosphate detection dye; absorbance at 650 nm is proportional to MurC activity, so inhibitors reduce the signal.
Specifications
| Assay Target | E. coli MurC activity |
|---|---|
| Target Type | Ligase |
| 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) |
| 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 MurC Assay (PDF, see Documents): Covers catalog # MURC100KE, MURC500KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli MurC 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.