E. coli Topoisomerase IV

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E. coli Topoisomerase IV

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Topo IV (from Escherichia coli) is prepared by overexpressing the parC and parE subunits in E. coli and purifying them by methods adapted from Peng and Marians, 19991.

The parC and parE 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
T4001 E. coli Topo IV
100 U
£127
T4002 E. coli Topo IV
500 U
£475
T4003 E. coli Topo IV
1000 U
£948
T4004 E. coli Topo IV
2000 U
£1,786
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E. coli Topoisomerase IV Relaxation Assay Kits

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These contain the enzyme 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
R4001 E. coli Topo IV Relaxation Kit 1
100 U of E. coli Topo IV and 50 µg of Supercoiled DNA
£210
R4002 E. coli Topo IV Relaxation Kit 2
500 U of E. coli Topo IV and 50 µg of Supercoiled DNA
£772
R4003 E. coli Topo IV Relaxation Kit 3
1000 U of E. coli Topo IV and 50 µg of Supercoiled DNA
£1,527
R4004 E. coli Topo IV Relaxation Kit 4
2000 U of E. coli Topo IV and 50 µg of Supercoiled DNA
£2,872
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E. coli Topoisomerase IV Decatenation Assay Kits

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These contain the enzyme 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
D4001 E. coli Topo IV Decatenation Kit 1
100 U of E. coli Topo IV and 20 µg kDNA
£210
D4002 E. coli Topo IV Decatenation Kit 2
500 U of E. coli Topo IV and 20 µg kDNA
£772
D4003 E. coli Topo IV Decatenation Kit 3
1000 U of E. coli Topo IV and 20 µg kDNA
£1,527
D4004 E. coli Topo IV Decatenation Kit 4
2000 U of E. coli Topo IV and 20 µg kDNA
£2,872
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E. coli Topoisomerase IV Assay Kits for Cell Extracts

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These kits are designed for assaying cell extracts and partially purified fractions containing over-expressed 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
TCE001 E. coli Topo IV Assay Kit 1 for Cell Extracts
For 100 assays
£130
TCE002 E. coli Topo IV Assay Kit 2 for Cell Extracts
For 500 assays
£383
TCE003 E. coli Topo IV Assay Kit 3 for Cell Extracts
For 1000 assays
£761
TCE004 E. coli Topo IV Assay Kit 4 for Cell Extracts
For 2000 assays
£1,454
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E. coli Topoisomerase IV Cleavage Assay Kits

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

Cleavage specific enzyme available separately on request.

Technical Documents

Cat No. Product Price Quantity
TCK001 E. coli Topo IV Cleavage Kit 1
100 U of E. coli Topo IV and 50 µg of supercoiled DNA
£380
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E.coli Topoisomerase IV ATPase kits

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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
ATPECT001 Kit for 100 ATPase assays
For determination of DNA-dependent ATPase activity; contains E. coli topoIV, buffers, ATP, PK-LDH, PEP, NADH and linear pBR322 DNA substrate.
£1,472
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High / Medium-Throughput Assay Kit - E. coli Topoisomerase IV

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The kit is supplied with sufficient E. coli 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
TRIV01 HTS Assay Kit - E. coli Topo IV 1-5 Kits
For 100 assays per plate
£420
TRIV02 HTS Assay Kit - E. coli Topo IV 6+ Kits
For 100 assays per plate
£330
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References

  1. Peng, H. and Marians, K.J. (1999) Overexpression and purification of bacterial topoisomerase IV, in DNA Topoisomerase Protocols Vol. I (Bjornsti, M-A., and Osheroff, N. eds.), Humana Press, Totowa, N.Jersey pp.163-169
  2. Maxwell, A., Burton, N.P. and O'Hagan, N. (2006) High-throughput assays for DNA gyrase and other topoisomerases. Nucleic Acid Res. 34 (15), e104