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Human topoisomerase II alpha is prepared by overexpressing in baculovirus-infected insect cells (Spodoptera frugiperda) and purifying it by methods developed in-house.
The enzyme is supplied in Dilution Buffer.
Store at -80°C.
It is recommended that larger pack sizes (500U and above) of the enzyme are aliquoted to avoid repeated freeze-thaw cycles.
See technical documents below for more detailed information and lot specific activities.
Human topoisomerase II beta is prepared by overexpressing in baculovirus-infected insect cells (Spodoptera frugiperda) and purifying it by methods developed in-house.
The enzyme is supplied at a concentration of 2-10 U/μl in Dilution Buffer.
Store at -80°C. (Stable for 3 months undiluted).
It is recommended that the enzyme is aliquoted to avoid repeated freeze-thaw cycles.
These contain human topo II and the catenated kDNA substrate in addition to the Assay and Dilution buffers for decatenation reactions. 1 U of human topo II 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. The kits are available with either the alpha or beta forms of the enzyme.
These contain human topo II and the supercoiled DNA substrate in addition to the Assay and Dilution buffers for relaxation reactions. 1 U of human topo II will relax 0.5 µg supercoiled pBR322 DNA in 30 minutes at 37°C. The kits are available with either the alpha or beta forms of the enzyme.
These kits are designed for assaying cell extracts and partially purified fractions containing human topo II. They contain kDNA substrate, Assay buffer, Dilution buffer, decatenated and linear DNA markers and stop buffer / loading dye.
The kit components are based on the standard assay which contains 500 ng substrate DNA per assay.
The kit is supplied with sufficient human topo II 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. The kits are available with either the alpha or beta forms of the enzyme.
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)
Human topoisomerase II alpha amino acids 1-453. This contains the ATPase domain, the activity of which can be stimulated by DNA (see Campbell and Maxwell (2002), J.Biol.Chem. 320 p.171. The protein is expressed in E.coli.
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