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Bacterial RNA polymerase makes the mRNA, tRNA and rRNA of the cell. The bacterial holoenzyme is built from several subunits with a total mass of about 400 kDa, and it is a target for the discovery of new antibacterial compounds.
Kit Components
| Component | PPA1000 |
|---|---|
| Buffer RP | 3200 µl |
| 100 x NTP mix | 310 µl |
| 100 x DNA | 310 µl |
| 1000 x Pyrophosphatase | 32 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 42 ml |
All reagents except the RNA polymerase are included.
Assay Principle
The reagent set follows RNA synthesis on a double-stranded DNA template by measuring the pyrophosphate released in the reaction. Pyrophosphatase in the reaction converts the pyrophosphate to phosphate, which is then read with a phosphate detection dye by absorbance at 650 nm; inhibitors of the polymerase lower the signal.
Specifications
| Assay Target | RNA polymerase activity (PPi) |
|---|---|
| Target Type | RNA Polymerase |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Colorimetric |
| Detection Wavelength | Absorbance 650 nm |
| Plate Format | 384-well plate |
| Instrument Required | Absorbance microplate reader |
| Assay Time | About 63 min (60 min reaction + 3 min detection) |
| Size | 1000 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- RNA Polymerase Pyrophosphate Assay (PDF, see Documents): Covers catalog # PPA1000.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this RNA polymerase activity (PPi) assay — including larger pack sizes, bulk reagents, 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.