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| Main SKU |
MurF carries out the last step in making UDP-MurNAc-pentapeptide in bacterial peptidoglycan biosynthesis. It adds the dipeptide D-Ala-D-Ala to UDP-MurNAc-tripeptide, coupled to ATP hydrolysis that releases ADP and inorganic phosphate. It is essential and an attractive antibacterial drug target.
Kit Components
| Component | MURF100KP |
|---|---|
| 10 x Buffer | 400 µl |
| 100 x UDP-MurNAc-tripeptide (UMAG-DAP) | 35 µl |
| 100 x D-Ala-D-Ala | 35 µl |
| 100 x ATP | 35 µl |
| 1000 x P. aeruginosa MurF | 10 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 5 ml |
Assay Principle
The kit measures the inorganic phosphate released when MurF adds D-Ala-D-Ala to UDP-MurNAc-tripeptide and hydrolyzes ATP with a phosphate detection dye; absorbance at 650 nm is proportional to MurF activity, so inhibitors reduce the signal.
Specifications
| Assay Target | P. aeruginosa MurF activity |
|---|---|
| Target Type | Ligase |
| Organism | P. aeruginosa |
| 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) |
| Size | 100 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- P. aeruginosa MurF Assay (PDF, see Documents): Covers catalog # MURF100KP.
Safety & Handling
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