Ogg1 Assay Kit 

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

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.

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.

sufficient for 180 assays

dry ice

−20°C

Zharkov, D.O., et al., Substrate specificity and reaction mechanism of murine 8-oxoguanine-DNA glycosylase. J. Biol. Chem. 275, 28607-28617 , (2000) abstract

UN 3316 9/PG 3

Ogg1 Substrate 8-oxo-G strand 1 vial

Ogg1 Substrate complementary strand 1 vial

Reaction buffer 10X 1.5 mL

Stop Solution 1mL

Ogg1 (control enzyme) 10 μg

Putrescine 1 mL

SigmaSpin Post-Reaction Purification Columns 2 each