| Source: |
Purified from E. coli containing a recombinant plasmid harboring the hOGG1 gene. |
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| Unit Definition: |
One Unit cleaves 1 pmole of a labeled oligonucleotide probe containing 8-oxoG base paired with C within a duplex oligo.
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| Assay Conditions: |
1X REC™ Buffer 6 (20 mM Tris-Cl (pH 8.0), 1 mM EDTA, 1
mM DTT, 100 µg/ml BSA), 4 pmoles of labeled 8-oxo-dG oligonucleotide annealed to the
compliment oligonucleotide, and serial dilutions of enzyme in a reaction volume of 20 μl are
incubated for 1 hour at 37°C. For analysis, 20 µl of 2X Loading Buffer (20 mM EDTA, 95%
formamide, and 0.13% bromophenol blue) are added, the samples are heated to 95°C for 10
min then fast cooled to 4°C, and the cleavage products are resolved by 20% denaturing
polyacrylamide gel electrophoresis, and percent cleavage quantified.
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| Applications: |
Detection of oxidative DNA damage
hOGG1 FLARE™ Assay
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| Storage: |
Store in working aliquots at -20∞C in a manual defrost freezer to avoid repeated freeze-thaws. The enzyme is supplied in 20 mM Tris-Cl (pH7.8), 1 mM EDTA, 100 mM NaCl, 1 mM DTT, 50% glycerol. |
