S. aureus Topoisomerase IV

Topo IV (from Staphylococcus aureus) is prepared by overexpressing the subunits in E. coliand purifying them by methods developed in-house.

The subunits are purified to >95% purity as judged by SDS-PAGE. The topo IV is supplied as a heterotetramer complex  in Dilution buffer.

It is recommended that the enzyme is aliquoted to avoid repeated freeze-thaw cycles. Store at -80ºC.

All enzyme is supplied with 5X concentrated Assay Buffer and Dilution buffers which are also available separately.

See technical documents below for more detailed information and lot specific activities.

Technical Documents

Cat No.	Product	Price	Quantity
SAT4001	S. aureus Topo IV100 U	£135
SAT4002	S. aureus Topo IV500 U	£499

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These contain S. aureus topo IV and the supercoiled DNA substrate in addition to the Assay and Dilution buffers for relaxation reactions. 1 U of topo IV will relax 0.5 µg supercoiled pBR322 DNA in 30 minutes at 37°C. 

Technical Documents

Cat No.	Product	Price	Quantity
SAR4001	S.aureus Topo IV RelaxationAssay Kit 1100 U of S. aureus Topo IV and 50 µg of Supercoiled DNA	£220
SAR4002	S.aureus Topo IV RelaxationAssay Kit 2500 U of S. aureus Topo IV and 50 µg of Supercoiled DNA	£816

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These contain S. aureus topo IV and the catenated kDNA substrate in addition to the Assay and Dilution buffers for decatenation reactions. 1 U of topo IV will decatenate 200 ng of kDNA when incubated in 1X Assay buffer in a total reaction volume of 30 µl at 37°C for 30 minutes. 

Technical Documents

Cat No.	Product	Price	Quantity
SAD4001	S.aureus Topo IV Decatenation Assay Kit 1100 U of S. aureus topo IV and 20 µg of kDNA	£220
SAD4002	S.aureus Topo IV Decatenation Assay Kit 2500 U of S. aureus topo IV and 100µg of kDNA	£816

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These kits are designed specifically for cleavage reactions. They contain S. aureus topo IV enzyme, supercoiled pBR322 DNA substrate and the Assay and Dilution buffers required for DNA cleavage reactions in addition to linearised pBR322 marker.

Cleavage specific enzyme available separately on request.

Technical Documents

Cat No.	Product	Price	Quantity
SATC001	S.aureus Topo IV Cleavage Assay Kit 1100 U of S. aureus topo IV and 50 ug of supercoiled DNA	£399

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These kits are designed for assaying cell extracts and partially purified fractions containing over-expressed S. aureus topo IV and contain supercoiled DNA substrate, Assay buffer, Dilution buffer, control relaxed DNA and stop buffer/loading dye.

Technical Documents

Cat No.	Product	Price	Quantity
SATE001	S.aureus Topo IV Cell Extract Assay Kit 1For 100 assays	£136
SATE002	S.aureus Topo IV Cell Extract Assay Kit 2For 500 assays	£408

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These kits can be used to test the effects of potential ATPase inhibitors. For example, the coumarin drugs such as novobiocin inhibit the action of topoisomerase IV by competitively inhibiting the hydrolysis of ATP thus preventing supercoiling.

These assays are microtitre plate-based and thus large numbers of compounds can be screened in a relatively short period of time. They also continuous assays which can provide more information than an end point assay.

Technical Documents

Cat No.	Product	Price	Quantity
ATPSAT001	Kit for 100 ATPase assaysFor determination of DNA-dependent ATPase activity; contains E. coli topoIV, buffers, ATP, PK-LDH, PEP, NADH and linear pBR322 DNA substrate.	£1,402

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The kit is supplied with sufficient S. aureus topo IV enzyme, plasmid DNA substrate, buffers and other assay components* for 100 assays. The enzyme is supplied at a concentration of 10 U/μl in Dilution Buffer. The kit is also supplied with sufficient wash buffers for one 96-well plate. These buffers are supplied as 20X concentrates and must be diluted with ultra pure water prior to use.

More information about this assay can be found on the "Services" page under "High/Medium Throughput Assay".

Kit issued with limited licence for individual use only. 

Patent held by Inspiralis Ltd., Norwich, Norfolk, UK. (Patent No. GB0424953.8, US7838230)

Technical Documents

Cat No.	Product	Price	Quantity
SATRIV01	HTS Assay Kit S. aureus Topo IV 1 - 5 KitsFor 100 assays per plate	£440
SATRIV02	HTS Assay Kit S. aureus Topo IV 6+ KitsFor 100 assays per plate	£346

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