S. aureus 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.  

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. 

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. Staphylococcus aureus Topo IV is prepared by overexpressing the subunits in E. coli and 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.

Store at -80ºC.

References

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