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| Main SKU |
Bacterial RNA polymerase synthesizes the cell's mRNA, tRNA and rRNA. The holoenzyme is made of several subunits, about 400 kDa in total, and is a target in the search for new antibacterial compounds.
Kit Components
| Component | RPAP100KE |
|---|---|
| Buffer RP | 400 µl |
| 100 x DNA | 33 µl |
| 100 x NTP mix | 33 µl |
| 1000 x Pyrophosphatase | 5 µl |
| 100 x E. coli RNA polymerase | 33 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 4500 µl |
All reagents, including the E. coli RNA polymerase, are included for 100 reactions in 384-well format.
Assay Principle
The kit follows E. coli RNA polymerase activity 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 | E. coli RNA polymerase activity |
|---|---|
| Target Type | RNA Polymerase |
| Organism | E. coli |
| 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 | 100 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 # RPAP100KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli RNA polymerase 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.