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inspiralis/E. coli Topoisomerase IV/For 500 assays/TCE002

inspiralis/E. coli Topoisomerase IV/For 500 assays/TCE002
  • inspiralis/E. coli Topoisomerase IV/For 500 assays/TCE002
商品介绍

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.ProductPriceQuantity
T4001E. coli Topo IV100 U£133
T4002E. coli Topo IV500 U£499
T4003E. coli Topo IV1000 U£995
T4004E. coli Topo IV2000 U£1,875
Custom Quote

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.ProductPriceQuantity
R4001E. coli Topo IV Relaxation Kit 1100 U of E. coli Topo IV and 50 µg of Supercoiled DNA£220
R4002E. coli Topo IV Relaxation Kit 2500 U of E. coli Topo IV and 50 µg of Supercoiled DNA£810
R4003E. coli Topo IV Relaxation Kit 31000 U of E. coli Topo IV and 50 µg of Supercoiled DNA£1,603
R4004E. coli Topo IV Relaxation Kit 42000 U of E. coli Topo IV and 50 µg of Supercoiled DNA£3,016
Custom Quote

E. coli Topoisomerase IV DecatenationAssay 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.ProductPriceQuantity
D4001E. coli Topo IV Decatenation Kit 1100 U of E. coli Topo IV and 20 µg kDNA£220
D4002E. coli Topo IV Decatenation Kit 2500 U of E. coli Topo IV and 100 µg kDNA£810
D4003E. coli Topo IV Decatenation Kit 31000 U of E. coli Topo IV and 200 µg kDNA£1,603
D4004E. coli Topo IV Decatenation Kit 42000 U of E. coli Topo IV and 400 µg kDNA£3,016
Custom Quote

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.ProductPriceQuantity
TCE001E. coli Topo IV Assay Kit 1 for Cell ExtractsFor 100 assays£136
TCE002E. coli Topo IV Assay Kit 2 for Cell ExtractsFor 500 assays£402
TCE003E. coli Topo IV Assay Kit 3 for Cell ExtractsFor 1000 assays£799
TCE004E. coli Topo IV Assay Kit 4 for Cell ExtractsFor 2000 assays£1,527
Custom Quote

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.ProductPriceQuantity
TCK001E. coli Topo IV Cleavage Kit 1100 U of E. coli Topo IV and 50 µg of supercoiled DNA£399
Custom Quote

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.ProductPriceQuantity
ATPECT001Kit 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.£1,472
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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.ProductPriceQuantity
TRIV01HTS Assay Kit - E. coli Topo IV 1-5 KitsFor 100 assays per plate£440
TRIV02HTS Assay Kit - E. coli Topo IV 6+ KitsFor 100 assays per plate£346
Custom Quote

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
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自1995年以来,我们一直为制药行业和学术界提供拓扑异构酶产品和服务,以协助抗感染和抗癌市场的研究。Inspiralis以前是John Innes Enterprises的公司,它是根据拓扑异构酶研究领域的全球领先者之一Tony Maxwell教授的研究建立的。

我们位于约翰·因内斯中心和东英吉利大学诺里奇研究园的学术研究中心附近的实验室。我们与麦克斯韦教授的学术研究小组保持着密切的联系,帮助我们保持在拓扑异构酶领域的最前沿。我们员工的经验以及与学术团体的联系使我们能够提供高水平的支持,以确保研究人员能够使用我们的产品获得最佳结果。生产我们的酶和底物的科学家也提供技术支持。

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