E. coli RNA Polymerase Pyrophosphate Assay Kit Plus

SKU:BHT16300107
Overview
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E. coli RNA Polymerase Assay Pyrophosphate Kit Plus, a colorimetric kit for E. coli RNA polymerase activity and inhibitor screening. Pyrophosphate released during RNA synthesis is turned into phosphate and read at 650 nm in 384-well plates. Complete kit for 100 assays with enzyme, pyrophosphatase and dye.
Assay Target E. coli RNA polymerase activity
Detection Method Colorimetric
Detection Wavelength Absorbance 650 nm
Plate Format 384-well plate
Assay Time About 63 min
Options selector
Catalog no. Size
RPAP100KE 100 assays
Available Options

Select the variant that best fits your experiment. Availability and lead time may vary by option.

  • Options: Size: 100 assays
  • Lead time: options listed in "Availability Content"; other statuses may take longer.
  • Storage: Please contact us to confirm storage conditions.
  • Shipping: Shipped frozen.
  • Upon receipt: store at recommended temperature as soon as possible.
  • Sales terms and conditions: Please review prior to ordering.
Field Specification
Organism E. coli
Applications
  • Enzyme Activity
  • Compound Profiling
Assay type
  • Enzyme Activity Assay
  • Inhibitor Screening Assay
Assay target E. coli RNA polymerase activity
Assay area Antibacterial Drug Discovery
Target type RNA Polymerase
Detection method
  • Colorimetric
Detection wavelength Absorbance 650 nm
Instrument required Absorbance microplate reader
Plate format 384-well plate
Assay time
  • About 63 min
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.
Catalog no. (Mfr.) RPAP100KE
Main SKU BHT16300107
Drug Target Assay Kit

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

For Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The E. coli RNA Polymerase Pyrophosphate Assay Kit Plus is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Please contact us to confirm storage conditions.
Q.What does this kit measure?
A.It measures E. coli RNA polymerase activity by detecting the pyrophosphate released during RNA synthesis on a DNA template. It is used to screen RNA polymerase inhibitors in 384-well plates.
Q.What is included in the kit?
A.The E. coli RNA Polymerase Pyrophosphate Assay Kit Plus (100 assays) includes 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) and phosphate detection dye (4500 µl).
Q.Is a protocol available?
A.Yes. The manufacturer's protocol (RNA Polymerase Pyrophosphate Assay, linked under Documents) covers catalog # RPAP100KE.
Q.How long does the assay take?
A.About 63 min (60 min reaction + 3 min detection). See the protocol for the exact steps.
Q.How should the kit be stored?
A.The E. coli RNA Polymerase Pyrophosphate Assay Kit Plus is shipped frozen. Please contact us for long-term storage conditions.

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.

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Experience the power of Celltrypse™, c-LEcta's innovative enzyme solution for gentle and efficient cell dissociation. Request your free sample and discover a superior alternative for your cell culture workflows.

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