96-Well S. aureus Topo IV DNA Decatenation Assay Kit Plus

SKU:BHT16300081
Overview
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Fluorescence-based 96-well kit for measuring S. aureus topoisomerase IV DNA decatenation and screening inhibitors. Decatenated DNA is separated on a TDD filter plate and read at Ex 485 nm / Em 535 nm. S. aureus Topo IV included; one 96-assay size. Needs a vacuum filtration setup.
Assay Target S. aureus Topo IV DNA decatenation
Detection Method Fluorescence
Detection Wavelength Ex 485 nm / Em 535 nm
Plate Format 96-well filter plate
Options selector
Catalog no. Size
SDD96KE 96 assays
Available Options

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

  • Options: Size: 96 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 S. aureus
Applications
  • Enzyme Activity
  • Compound Profiling
Assay type
  • Enzyme Activity Assay
  • Inhibitor Screening Assay
Assay target S. aureus Topo IV DNA decatenation
Assay area Antibacterial Drug Discovery
Target type DNA Topoisomerase IV
Detection method
  • Fluorescence
Detection wavelength Ex 485 nm / Em 535 nm
Instrument required Fluorescence reader + vacuum device
Plate format 96-well filter plate
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.
Catalog no. (Mfr.) SDD96KE
Main SKU BHT16300081
Drug Target Assay Kit

DNA decatenation is an essential step of DNA replication in cells. In bacteria, topoisomerase IV (the parC-parE complex) carries out this reaction, converting concatenated DNA into decatenated DNA.

Kit Components

Component SDD96KE
10 x Assay buffer 600 µl
10 x Concatenated DNA 500 µl
10 mM ATP 110 µl
1000 x S. aureus topo IV 8 µl
Stop solution (0.4 M EDTA) 600 µl
20 x Fluorescence dye 260 µl
10 x Rinse buffer 2 ml
V-bottom plate 1 plate
TDD filter plate 1 plate
Black 96-well plate 1 plate

Final reaction conditions: 20 mM Tris-HCl pH 8, 35 mM NH4OAc, 4.6% glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 3 µg/ml concatenated DNA, 0.2 mM ATP and 5 nM S. aureus topoisomerase IV. A reaction without enzyme or ATP can serve as negative control.

Assay Principle

After the reaction, the sample is filtered through the TDD filter plate, which separates decatenated DNA from concatenated DNA. The decatenated DNA passes into a black 96-well plate, is stained with fluorescence dye and read at 535 nm (excitation 485 nm), so inhibitors lower the signal.

Specifications

Assay Target S. aureus Topo IV DNA decatenation
Target Type DNA Topoisomerase IV
Organism S. aureus
Assay Type Enzyme Activity Assay; Inhibitor Screening Assay
Detection Method Fluorescence
Detection Wavelength Ex 485 nm / Em 535 nm
Plate Format 96-well filter plate
Instrument Required Fluorescence reader + vacuum device
Equipment Needed Lab vacuum line or pump (up to 80 kPa or 600 mmHg), plate vacuum device, plate fluorescence reader (Ex 485 nm / Em 535 nm)
Size 96 assays
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.

Protocol

The manufacturer's protocol describes the procedure and reaction volumes.

  • 96-Well S. aureus Topo IV DNA Decatenation Assay (PDF, see Documents): Covers catalog # SDD96KE.

Safety & Handling

For Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The 96-Well S. aureus Topo IV DNA Decatenation 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 the DNA decatenation activity of S. aureus topoisomerase IV, which converts concatenated DNA into decatenated DNA. It is used for high-throughput testing of topoisomerase IV inhibitors.
Q.What is included in the kit?
A.The 96-Well S. aureus Topo IV DNA Decatenation Assay Kit Plus (96 assays) includes 10 x assay buffer (600 µl), 10 x concatenated DNA (500 µl), 10 mM ATP (110 µl), 1000 x S. aureus topo IV (8 µl), stop solution (0.4 M EDTA) (600 µl), 20 x fluorescence dye (260 µl), 10 x rinse buffer (2 ml), V-bottom plate (1 plate), TDD filter plate (1 plate) and black 96-well plate (1 plate).
Q.Is a protocol available?
A.Yes. The manufacturer's protocol (96-Well S. aureus Topo IV DNA Decatenation Assay, linked under Documents) covers catalog # SDD96KE.
Q.How should the kit be stored?
A.The 96-Well S. aureus Topo IV DNA Decatenation Assay Kit Plus is shipped frozen. Please contact us for long-term storage conditions.

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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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