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Human Topoisomerase I is prepared by overexpressing the enzyme in baculovirus-infected insect cells (Spodoptera frugiperda) and purifying it by methods adapted from Stewart et al., 1996. The enzyme is supplied μl in Dilution Buffer.
Store at -80°C. It is recommended that the enzyme is aliquoted to avoid repeated freeze-thaw cycles.
See technical documents below for more detailed information and lot specific activities.
These contain the enzyme and the supercoiled DNA substrate in addition to the Assay and Dilution buffers for relaxation reactions. 1 U of human topo I will relax 0.5 µg supercoiled pBR322 DNA in 30 minutes at 37°C.
These kits are designed for assaying cell extracts and partially purified fractions containing human topo I. They contain supercoiled DNA substrate, Assay buffer, Dilution buffer and relaxed DNA marker.
The kit components are based on the standard human topo I assay whch contains 500 ng substrate DNA per assay.
The kit is supplied with sufficient human topo I 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).