E. coli Topo IV DNA Relaxation Assay Kit Plus (enzyme included)

Mfr.No.	T4RA-100KE
Description	DNA topoisomerase IV (the parC-parE complex) have DNA relaxation activity that converts the supercoiled DNA into its relaxed form. The Topoisomerase IV DNA Relaxation Assay Kit is based on the principle that the relaxed DNA suppresses the fluorescent intensity much more than the supercoiled DNA when the DNA interacts with fluorescence dye H19 (a fluorescence dye for DNA relaxation / supercoiling assay) in the presence of magnesium. When the supercoiled DNA is converted into its relaxed form, the fluorescent signal decreases. The E. coli Topo IV DNA Relaxation Assay Kit Plus (T4RA-100KE) includes 480 µl of 10 x Buffer, 405 µl of 10 x supercoiled plasmid DNA, 45 µl of 100 x E. coli topoisomerase IV, 420 µl of 10mM ATP and 20 µl of 1500 x fluorescence dye H19 for 100 assays of DNA relaxation reactions in a 96-well assay format.
Drug target assays	Antibacterial drug targets

Product Manual	Datasheet
Protocols Description	Assay Protocol 1. Reaction and sample preparation: The total volume of each reaction mixture is 40 µl including: 24 µl of H2O, 4 µl of 10 x Buffer, 4 µl of 10 x supercoiled DNA, 4 µl of 10 mM ATP, 4 µl of 1000 nM Topoisomerase VI. Incubate the reaction mixture at 37°C for 60 min. The final concentrations are 20 mM Tris-HCl, pH 8, 35 mM NH4OAc, 4.6 % glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2 , 25 µg/ml supercoiled plasmid DNA, 1 mM ATP and 100 nM topoisomerase IV. 2. Assay (1) Make 1x H19 Dilution Buffer by dilution of the 10 x H19 Dilution Buffer 10-fold with water. Freshly prepare the H19 dye by dilution of 1 µl of the 1500 x H19 dye stock solution with 1500 µl of 1x H19 Dilution Buffer (1500 x dilution). Mix the solution evenly by inverting the tube a few times. (2) Mix 250 µl of the freshly prepared H19 dye with each reaction solution. Incubate the mixture for 5 min. (3) Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.

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Combination SKU	Supplier No.	Specifications	Your price
BHE16300118-1	T4RA-100KE	E. coli Topo IV	$737.30	Add to Cart
BHE16300118-2	T4RA-100KP	P. aeruginosa Topo IV	$737.30	Add to Cart
BHE16300118-3	T4RA-100KS	S. aureus Topo IV	$737.30	Add to Cart
BHE16300118-4	T4RA-100KN	S. pneumoniae Topo IV	$737.30	Add to Cart