T7428
Topoisomerase I from wheat germ
buffered aqueous glycerol solution
Synonym(s):
DNA relaxing enzyme, Topo I
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About This Item
grade
Molecular Biology
form
buffered aqueous glycerol solution
mol wt
97 kDa
foreign activity
RNase and DNase, none detected
shipped in
dry ice
storage temp.
−70°C
General description
Topoisomerase I relaxes supercoiled DNA molecules. The enzyme initiates transient breakages and rejoins of phosphodiester bonds in superhelical turns of closedcircular dsDNA.
Application
Suitable for:
- Analsis of degree of DNA supercoiling
- Forming knots and circles in single stranded DNA
- Converting complementary single stranded DNA to double stranded, circular forms
- Rejoining nicks in double-stranded DNA
Biochem/physiol Actions
Topoisomerase I relaxes supercoiled DNA molecules. The enzyme initiates transient breakages and rejoins of phosphodiester bonds in superhelical turns of closed-circular DNA. Enzyme activity is independent of right- and left-handed superhelices.
Other Notes
One unit will convert 1μg of supercoiled pGEM9Zf(−) plasmid DNA to the relaxed form in 30 min at 37 °C.
To preserve enzyme activity, avoid multiple freeze-thaw cycles and frequent temperature variations.
Topoisomerase I is supplied in a solution of 50 mM Tris-HCl, pH 7.9, 1 mM EDTA, 1 mM DTT, 0.5 M NaCl and 50% glycerol (v/v).
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
Regulatory Information
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Related Content
Instructions
J C Wang
Annual review of biochemistry, 65, 635-692 (1996-01-01)
The various problems of disentangling DNA strands or duplexes in a cell are all rooted in the double-helical structure of DNA. Three distinct subfamilies of enzymes, known as the DNA topoisomerases, have evolved to solve these problems. This review focuses
P D Foglesong et al.
BioTechniques, 13(3), 402-404 (1992-09-01)
Supercoiled and relaxed DNA were resolved electrophoretically in the presence of 0.5 micrograms/ml ethidium bromide. Under these conditions the Gaussian distributions of topological isomers of both supercoiled and relaxed DNA migrated as discrete bands. The separation of these DNAs was
Maris A Cinelli et al.
Journal of medicinal chemistry, 55(24), 10844-10862 (2012-12-12)
Hydroxylated analogues of the anticancer topoisomerase I (Top1) inhibitors indotecan (LMP400) and indimitecan (LMP776) have been prepared because (1) a variety of potent Top1 poisons are known that contain strategically placed hydroxyl groups, which provides a clear rationale for incorporating