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About This Item
UNSPSC Code:
12352200
usage
sufficient for 180 assays
shipped in
dry ice
storage temp.
−20°C
General description
The assay is based on Ogg1 glycosylase activity that recognizes and removes the mutated base (8-oxo-G). The substrate is a 23 oligonucleotide containing 8-oxo-dG at its 11th base, labeled with 32P at its 5′ end, and annealed to its complementary strand (containing dC at the opposite base position to the 8-oxo-dG). Upon cleavage of the substrate by the Ogg1 enzyme, the oligonucleotide strands are run on a denaturing gel and a 10 base fragment (labeled cleavage product) is revealed in addition to the original 23 base oligonucleotide band. The detection is performed by autoradiography.
Biochem/physiol Actions
Ogg1, a DNA repair protein, is involved in the repair of the major product of DNA oxidation, the miscoding base 8-oxoguanine (8-oxo-G). Polymorphism in the human OGG1 gene is associated with the risk of various cancers such as lung and prostate cancer.
Features and Benefits
Ogg1 kit
- provides a tool for measurement of Ogg1 enzyme activity
- an Ogg1 control enzyme enables screening of Ogg1 modulators
- can be used with cell lysates and purified enzyme preparation
Kit Components Only
Product No.
Description
- Ogg1 Substrate 8-oxo-G strand 1 mL/vial
- Ogg1 Substrate complementary strand 1 mL/vial
- Reaction buffer 10X 1.5 mL
- Stop Solution 1 mL
- Ogg1 (control enzyme) 10 μg
- Putrescine 1 mL
- SigmaSpin Post-Reaction Purification Columns 2 ea
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Instructions
D O Zharkov et al.
The Journal of biological chemistry, 275(37), 28607-28617 (2000-07-08)
Genomic DNA is prone to oxidation by reactive oxygen species. A major product of DNA oxidation is the miscoding base 8-oxoguanine (8-oxoG). The mutagenic effects of 8-oxoG in mammalian cells are prevented by a DNA repair system consisting of 8-oxoguanine-DNA