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Topoisomerase DNA cleavage Assay Kit
제품명: Topoisomerase DNA cleavage Assay Kit
용도: Drug target and enzyme assays
메이커: ProFoldin
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소개

 

 

Topoisomerase DNA cleavage Assay Kit 

 

Drug target and enzyme assays

 

 


 

Bacterial gyrase, topoisomerase IV (parC-parE) and mammalian topoisomerase II are type II topoisomerases. These topoisomerases change DNA topology by cleavage and religation of both DNA strands. When an inhibitor such as a fluoroquinolone drug blocks the religation step of the topoisomerase reaction, the covalent complex between the cleaved or linearized DNA and the topoisomerase (cleavage complex) is accumulated. Formation of the DNA cleavage complex causes DNA damage in the cells (topoisomerase poison). Testing formation of DNA cleavage complexes is useful for understanding the topoisomerase inhibition mechanism of novel topoisomerase inhibitors. 


The 96-well topoisomerase DNA cleavage assay kits are based on the principle that the linearized DNA has much greater permeability through a solution matrix and a filter than the closed circular DNA. For example, both novobiocin and ciprofloxacin inhibit DNA gyrase. However, novobiocin does not cause formation of cleavage complex. Ciprofloxacin does. 


The assay kit is in 96-well plate format and can be used for testing topoisomerase poison compounds in a high throughput setting.

 

 

 

 

Topoisomerase DNA cleavage assays.PNG

 

 

 

 

 

96-well E. coli Gyrase DNA Cleavage Assay Kit Plus (Catalog No. T2C96KE)

Kit components: 300μl 10 x TDC buffer, 30μl 100 x DNA, 260μl SDS solution, 260μl Protease K, 10 ml TDC matrix, 6 ml Rinse buffer, 500μl 10 x TDC dye, 1 TDC filter plate, 30μl 100 mM ATP, 1 black plate, 1 V-bottom plate, 26μl 100 x E. coli gyrase. 


It is for 96 assays in a 96-well plate format.


 


 

96-well Topoisomerase DNA Cleavage Assay Kit (enzyme not included) Catalog No. TDC96K)

Kit components: 300μl 10 x TDC buffer, 30μl 100 x DNA, 260μl SDS solution, 260μl Protease K, 10 ml TDC matrix, 6 ml Rinse buffer, 500μl 10 x TDC dye, 1 TDC filter plate, 30μl 100 mM ATP, 1 black plate, 1 V-bottom plate. 


It is for 96 assays in a 96-well plate format.


 


 

96-well E. coli Topo IV DNA Cleavage Assay Kit Plus (Catalog No. T4C96KE)


Kit components: 300μl 10 x TDC buffer, 30μl 100 x DNA, 260μl SDS solution, 260μl Protease K, 10 ml TDC matrix, 6 ml Rinse buffer, 500μl 10 x TDC dye, 1 TDC filter plate, 30μl 100 mM ATP, 1 black plate, 1 V-bottom plate, 26μl 100 x E. coli topoisomerase IV. 


It is for 96 assays in a 96-well plate format.

 



 

96-well S. aureus Topo IV DNA Cleavage Assay Kit Plus (Catalog No. T4C96KS)

Kit components: 300μl 10 x TDC buffer, 30μl 100 x DNA, 260μl SDS solution, 260μl Protease K, 10 ml TDC matrix, 6 ml Rinse buffer, 500μl 10 x TDC dye, 1 TDC filter plate, 30μl 100 mM ATP, 1 black plate, 1 V-bottom plate, 26μl 100 x S. aureus topoisomerase IV. 


It is for 96 assays in a 96-well plate format.