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Nicotinate phosphoribosyltransferase (NAPRTase, PncB) is part of the NAD salvage pathway. It makes nicotinic acid mononucleotide (NAMN) by transferring the phosphoribosyl group of phosphoribosyl pyrophosphate (PRPP) to nicotinate (NA), releasing pyrophosphate. PncB is a drug target in Gram-positive bacteria.
Pack Sizes
Both sizes include S. aureus PncB and pyrophosphatase. The only difference is the number of assays: PNCB100KS provides reagents for 100 assays and PNCB500KS for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (PNCB100KS) | 100 assays; enzyme included |
|---|---|
| 500 assays (PNCB500KS) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (PNCB100KS) | 500 assays (PNCB500KS) |
|---|---|---|
| 10 x Buffer | 400 µl | 2 ml |
| 100 x NA (nicotinate) | 35 µl | 170 µl |
| 100 x PRPP | 35 µl | 170 µl |
| 100 x ATP | 35 µl | 170 µl |
| 100 x S. aureus PncB (5000 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 |
Assay Principle
Pyrophosphate released by PncB is converted into inorganic phosphate by the included pyrophosphatase, and the phosphate is detected with a dye by absorbance at 650 nm. Reactions can run in 384-well or 96-well plates, or in tubes with a cuvette readout.
Specifications
| Assay Target | S. aureus PncB activity |
|---|---|
| Target Type | Phosphoribosyltransferase |
| Organism | S. aureus |
| 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.
- S. aureus PncB Assay (PDF, see Documents): Covers catalog # PNCB100KS, PNCB500KS.
Safety & Handling
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