Dna relaxation assay
WebCross contamination with topo I was assessed by assaying for relaxation of supercoiled DNA under conditions where Human Topoisomerase II Alpha activity is suppressed. Excess purified enzyme was incubated with pRYG DNA in the absence of ATP (with and without Mg++) for 2 hr at 37° C. WebDecatenation, rather than relaxation, is the preferred reaction for topo IV and human topo II making this an ideal assay for the quick identification of potential inhibitors. The IC 50 for inhibition of decatenation can be visually assessed as the concentration of compound which leads to a 50% reduction in the mini-circle band.
Dna relaxation assay
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WebJun 1, 2012 · The protocols described in this unit are for assays used to assess new chemical entities for their ability to inhibit both forms of DNA topoisomerase. Included are an in vitro assay for... WebFeb 8, 2024 · For the DNA relaxation assay, pUC19 supercoiled plasmid (300 ng) was used as the substrate and the assay was performed without ATP in the reaction. Ciprofloxacin at 25 µM and novobiocin at 20...
WebFig. 1 Superhelical DNA relaxation properties of the quinolone resistant ParC S80L Topo IV. A, standard superhelical DNA relaxation assays containing 10 n M DNA and the indicated amounts of either wild-type or ParC S80L Topo IV were incubated, processed, and analyzed as described under “Materials and Methods.” WebIt is for 100 DNA relaxation assays in a 96-well plate format or 200 assays in a 384-well assay format. Human topo I enzyme in not included and can be purchased separately. …
WebMay 13, 2008 · Therefore, the relaxation assay (described in Materials and Methods) monitors the fate of the DNA during single steps of phosphodiester synthesis, with … WebAug 10, 2024 · Using a sterile loop, inoculate the DNase agar with the organism to be tested on the test area. Incubate the plate at 35-37°C for 24 hours. After incubation observe the color change in DNase with methyl …
WebJan 1, 2001 · This is a convenient assay for comparing the inhibitory potencies of series of drugs. 52 Conclusion The relaxation and cleavage assays presented here are the most …
WebThese 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 which contains 500 ng substrate DNA per assay. Technical Documents display schermo lcd lg 406WebProduct Description includes 160 ul of 10 x Buffer HT1, 84 ul of 10 x supercoiled plasmid DNA, 8 ul of 1500 x Dye H19 and 500 ul of 10 x H19 dilution buffer. It is for 20 DNA relaxation assays in a 96-well plate format or 40 assays in a 384-well assay format. Human to Specifications Inventory Number: PTFHRA020KKT Part Number: HRA020K … cpk and ppk minitabWebDNA relaxation assay revealed that compound 22 showed potent inhibitory activity against Topo IIα in vitro. Compound 22 also induced DNA breaks in MDA-MB-435 cells evidenced by comet tails and the accumulation of γ-H2AX foci. The ability of 22 in inducing DNA breaks mediated by Topo IIα resulted in G2/M phase arrest and apoptosis. Conclusion: cpk anestesiaWebJan 13, 2016 · Topo II mediated DNA relaxation assay results showed that derivatives could significantly inhibit the activity of Topo II, and the structure–activity relationship studies indicated the importance of the alkylamino side chain and the benzoazolyl group. cpk and process capabilityWebFeb 1, 2024 · DNA Relaxation Assays. The human TOP2A and TOP2B were used to perform DNA relaxation assays. TOP2 enzyme was incubated with the supercoiled … cpk and six sigmaWebOct 1, 2024 · Topoisomerase assays. DNA topoisomerase enzymes were supplied by Inspiralis Ltd, and assays were carried out following the manufacturer's protocols. ... Further addition of topo IIα in excess of 130 … cpk archivesWebThe supercoiled DNA relaxation assay results for Top68W/A suggested that the loss of the Van der Waals and electrostatic interactions made by Trp 203/205/206, due to their mutation to Ala ... cpka pharma conference 2023