DNA Unwinding Assay Kit

This kit is designed to investigate whether a compound intercalates into the DNA double-helix, or binds in the groove, leading to unwinding of the DNA. This is a characteristic of a number of inhibitors of DNA-modifying enzymes such as the topoisomerase inhibitor m-amsacrine.

Intercalators contain planar, normally polycyclic, aromatic structures which can insert between the bases of the double-helical DNA molecule. Compounds which are able to intercalate into DNA or bind in the groove can lead to local unwinding of the DNA leading to a decrease in the twist of the DNA. If a DNA molecule is nicked and rejoined (for example by a topoisomerase) in the presence of such a compound, then the result is relaxed, underwound DNA; on removal of the enzyme and compound this converts to supercoiled DNA. The supercoiled DNA formed in these conditions is indicative of an intercalator.

This is the basis of the DNA unwinding assay. Supercoiled (or relaxed) plasmid DNA is incubated briefly with the test compound prior to relaxation by the wheat germ topo I. The enzyme and drug are then removed by extraction and the plasmid analysed by gel electrophoresis. Supercoiled topoiosmers indicate the compound is an intercalator or groove binder. If the compound is not an intercalator then the product will be relaxed plasmid.

The assay should be checked by performing a control reaction using relaxed plasmid to show that the compound is not simply acting as an inhibitor of the topo I.

The kit (product code DUKSR001) contains all you need to do the assay including:-

Supercoiled pBR322 (50 μg), relaxed pBR322 (25 μg), wheat germ topo I (250 U), Assay Buffer (1 ml) and Dilution Buffer (1 ml).

We also supply kits which contain topo I and buffers but only:-

(i) the relaxed form of pBR322 (product code DUKR002). This will show if a compound is an intercalator but not if it also inhibits the topo I (i.e. false-negatives are possible)

(ii) the supercoiled form of pBR322 (product code DUKS003). This will show if a compound is an intercalator but the same result will be given if it is an inhibitor of topo I (i.e. false-positives are possible)

This is illustrated in the scheme below:

unwinding figure

Technical Documents

Cat No.	Product	Price
DUKSR001	DNA Unwinding Assay Kit100 assays	£268
DUKR002	DNA Unwinding Assay Kit - Relaxed only100 assays	£204
DUKS003	DNA Unwinding Assay Kit - Supercoiled only100 assays	£204

Custom Quote

品牌介绍

Inspiralis

Inspiralis在提供高质量拓扑异构酶，连接酶和解旋酶方面享誉全球，包括检测试剂盒，底物和筛选服务。

We are located in laboratories adjacent to the academic research centres of the John Innes Centre and the University of East Anglia on the Norwich Research Park. We maintain close links with the academic research group of Professor Maxwell, helping us to remain at the forefront of the topoisomerase field. The experience of our staff and the connections with the academic group allow us to provide a high level of support to ensure that researchers are able to get the best results using our products. The same scientists who produce our enzymes and substrates also provide the technical support.

Our aim is to provide pharmaceutical companies, and others involved in drug development, with the necessary tools to develop and screen novel anti-infective and anti-cancer compounds. In order to achieve this we will continue to expand the range of topoisomerase products and screening services we offer.

产品列表：

No.	品牌	货号	名称	规格
1	Inspiralis	7F001	7F11 Monoclonal	0.1ml
2	Inspiralis	PA001	Anti-Gyr A Polyclonal	0.1 ml
3	Inspiralis	HT001	Anti-human topo II polyclonal - 0.1mL	0.1 ml
4	Inspiralis	G1001	E.coli gyrase	100 U
5	Inspiralis	T4001	E. coli Topo IV	100 U
6	Inspiralis	SAG4001	S. aureus gyrase	100 U
7	Inspiralis	SAT4001	S. aureus Topo IV	100 U
8	Inspiralis	MTG001	M. tuberculosis Gyrase (HIS)	100 U
9	Inspiralis	MTT01	M.tuberculosis Topo I	100 U
10	Inspiralis	PAG1001	P. aeruginosa Gyrase	100 U
11	Inspiralis	PAT4001	P. aeruginosa Topo IV	100 U
12	Inspiralis	SPG1001	S. pneumoniae Gyrase	100 U
13	Inspiralis	SPT4001	S. pneumoniae Topo IV	100 U
14	Inspiralis	ABG1001	A. baumannii Gyrase	100 U
15	Inspiralis	ABT4001	A. baumannii Topo IV	100 U