High / Medium-Throughput Topoisomerase Assay

A high-throughput microtitre plate-based assay was developed in the laboratory of Professor A. Maxwell of the John Innes Centre along with staff of Inspiralis. It is based upon the fact that negatively-supercoiled plasmids form intermolecular triplex DNA more readily than do relaxed plasmids. This assay overcomes some of the problems of gel-based assays, which are time consuming and are therefore inherently low-throughput. Inspiralis are able to offer this assay as a means of screening compounds for inhibitory activity against topoisomerases including gyrase, topo IV and human topos I and II. The assay is adaptable to other enzymes that change DNA supercoiling such as restriction enzymes. Some of the results of a comparison of the assay with the conventional gel-based assay are shown below.

The assay is particularly useful for screening of compounds and may also be used for determination of IC50 values. In the examples given below, the determination of IC50values for the inhibition of supercoiling by gyrase and relaxation by topo IV and human topo I and II by the plate- and gel-based assays are compared. Similar inhibition profiles and IC50 values were obtained in both assays.

graphs gyrasegraphs topo ivgraphs human

Pricing

The prices for this assay are as follows:

  • One off set up fee for either fixed concentration of inhibitor or IC50 determination £500.00
  • For inhibition at a fixed concentration for up to 40 compounds in duplicate with controls £400.00
  • For additional compounds above 40 £5.00 per cpd
  • For IC50 determinations for up to 5 compounds in duplicate with controls £400.00
  • For additional compounds above 5 £80.00 per cpd

Contact us now to discuss your requirements: contact@inspiralis.co.uk.

The assay is also available in kit form. See individual enzymes in Products section for details.