E. coli Topoisomerase IV Decatenation Assay Kits

These kits contain all the components necessary to perform decatenation reactions. Including the enzyme, the catenated BisCat DNA or kDNA substrate, a decatenated kDNA marker (for kDNA kits), the Assay and Dilution buffers as well as 10X GSTEB - our new stop and loading buffer.

Our NEW BisCat DNA decatenation substrate consists of two supercoiled, singly-linked minicircles of 2.3 and 2.6 kb. Type II topoisomerases can decatenate the substrate to produce two supercoiled minicircle products.

DNA Topoisomerase IV (Topo IV) is a bacterial type II topoisomerase which can decatenate and relax supercoiled DNA and it is a validated target for antibacterial compounds, such as the quinolones and the coumarins. Escherichia coli Topo IV is prepared by overexpressing the ParC and ParE subunits in E. coli and purifying them by methods adapted from Peng and Marians, 1999¹.

1 U of topo IV will decatenate 200 ng of BisCat DNA or kDNA when incubated in 1X Assay buffer in a total reaction volume of 30 µl at 37°C for 30 minutes.

Cat no.	Product	Price
DB4001	E. coli Topoisomerase IV BisCat Decatenation Kit 1 100 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 20 µg BisCat DNA, and 10X GSTEB	£563.00
D4001	E. coli Topoisomerase IV kDNA Decatenation Kit 1 100 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 20 µg kDNA, decatenated kDNA marker and 10X GSTEB	£521.00
DB4002	E. coli Topoisomerase IV BisCat Decatenation Kit 2 500 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 100 µg BisCat DNA, and 10X GSTEB	£2,064.00
D4002	E. coli Topoisomerase IV kDNA Decatenation Kit 2 500 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 100 µg kDNA, decatenated kDNA marker and 10X GSTEB	£1,941.00
DB4003	E. coli Topo IV BisCat Decatenation Assay Kit 3 1000 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 200 µg BisCat DNA, and 10X GSTEB	£3,457.00
D4003	E. coli Topoisomerase IV kDNA Decatenation Kit 3 1000 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 200 µg kDNA, decatenated kDNA marker and 10X GSTEB	£3,395.00
DB4004	E. coli Topo IV BisCat Decatenation Assay Kit 4 2000 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 400 µg BisCat DNA, and 10X GSTEB	£5,852.00
D4004	E. coli Topoisomerase IV kDNA Decatenation Kit 4 2000 U of E. coli Topo IV supplied with 5X Assay Buffer, Dilution Buffer, 400 µg kDNA, decatenated kDNA marker and 10X GSTEB	£5,626.00

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References

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
Waraich NF, Jain S, Colloms SD, Stark WM, Burton NP, Maxwell A. A novel decatenation assay for DNA topoisomerases using a singly-linked catenated substrate. Biotechniques. 2020 Nov;69(5):356-362