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Bacterial RNA polymerase makes the mRNA, tRNA and rRNA of the cell. The holoenzyme is built from several subunits, and blocking it is a route to new antibacterial compounds.
Kit vs Kit Plus
Both formats contain the same buffer, DNA template, NTPs and dye for 100 assays. The only difference is that the Kit Plus (RPA100KE) also includes 33 µl of 100 x E. coli RNA polymerase.
| Kit (enzyme not included), 100 assays (RPA100K) | 100 assays |
|---|---|
| Kit Plus (enzyme included), 100 assays (RPA100KE) | 100 assays |
Kit Components
| Component | Kit (enzyme not included), 100 assays (RPA100K) | Kit Plus (enzyme included), 100 assays (RPA100KE) |
|---|---|---|
| 10 x Buffer | 400 µl | 400 µl |
| 100 x DNA | 33 µl | 33 µl |
| 100 x NTPs | 33 µl | 33 µl |
| 10 x Fluorescence dye | 350 µl | 350 µl |
| 100 x E. coli RNA polymerase | Not included | 33 µl |
The 100 x NTP mix is 50 mM.
Assay Principle
The RNA polymerase copies a DNA template into RNA using the NTP mix. A fluorescence dye that binds the newly made RNA is then added, and the signal at 535 nm (excitation 485 nm) rises with polymerase activity; inhibitors 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 | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | 384-well or 96-well plate |
| Instrument Required | Fluorescence microplate reader |
| 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.
- E. coli RNA polymerase Assay (PDF, see Documents): Covers catalog # RPA100K, RPA100KE.
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.