Gel-based E. coli DNA Topoisomerase II (Gyrase) Assay Kit Plus-100 (for 100 assays)

Mfr.No.	GDSA100KE
Description	DNA topoisomerases such as bacterial topoisomerase II (gyrase) convert relaxed circular DNA into supercoiled DNA (DNA supercoiling reaction). The Gel-Based E. coli Topoisomerase II (Gyrase) DNA Supercoiling Assay Kit is based on the principle that the supercoiled DNA and relaxed DNA are separated by agarose gel electrophoresis. Fluorescence-based DNA supercoiling assays in a 96-well pate format are also available for high throughput screening of gyrase inhibitors. For more information of the high throughput gyrase DNA supercoiling assays, please visit the website at http://www.profoldin.com/topoisomerase_assays_1.html. The Gel-Based E. coli DNA Topoisomerase II (Gyrase) Assay Kit Plus-100 (Catalog No. GDSA100KE) includes all the reagents for 100 samples in gel-based assays of E. coli gyrase DNA supercoiling activity. It includes 400 µl of 10 x Buffer, 105 µl of 250µg/ml relaxed DNA, 220 µl of 10 mM ATP, 550 µl of 5 x Gel loading buffer and 25 µl of 2000 nM E.coli gyrase (100 x).
Drug target assays	Antibacterial drug targets

Product Manual	Datasheet
Protocols Description	Assay Protocol 1. Reaction: The total volume of each reaction mixture is 20µl including: 13µl of H2O, 2µl of 10 x Buffer, 1µl of 250µg/ml relaxed DNA, 2µl of 200 nM E. coli gyrase (10 x), 2µl of 10 mM ATP. Incubate the reaction mixture at 37°C for 60 min. At the end of the reaction, add 5µl of 5 x gel loading buffer. Note: 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, 12.5µg/ml relaxed plasmid DNA, 1 mM ATP and 20 nM topoisomerase II. The 10 x enzyme is prepared by dilution of the 100 x enzyme in 1x assay buffer. A negative control reaction can be the reaction mixture without addition of ATP. 2. Agarose gel electrophoresis (1) Prepare 1 % agarose gel in 1 x TAE buffer. (2) Load 25 ul of the sample. (3) Run the gel at 100 V for 90 min to 2 hours. (4) Stain the gel in an ethidium bromide solution and destain the gel in water. (5) Take a picture of the gel under UV light.

Have you used this product?

SKU	Supplier No.	Size	Your price	Quantity
BHE16300100	GDSA100KE	100 assays	$499.10		Add to Cart