| Source: |
Purified from E. coli containing a recombinant plasmid harboring the mouse Aag protein. |
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| Unit Definition: |
One unit cleaves 1 pmol of a 32P-labeled oligonucleotide probe containing hypoxanthine within an oligonucleotide duplex in one hour at 37°C. |
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| Assay Conditions: |
1X RECTM Buffer 9 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 1 mM EDTA, 1 mM EGTA, and 0.1 mM DTT), 4 pmoles 32P hypoxanthine oligonucleotide, 4 pmoles complementary oligonucleotide, and serial dilutions of enzyme in a reaction volume of 20 µl. Incubate for 1 hour at 37°C. Reaction products are resolved by 20% denaturing PAGE. |
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| Applications: |
CometAssay™/FLARE™
Supercoiled relaxation assays
DNA fragmentation analysis
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| Storage: |
Store at -20°C. For long-term storage freeze in working aliquots at -80°C to avoid repeated freeze-thawing. The enzyme is stored in 10 mM HEPES-KOH (pH 7.4); 100 mM KCl; 1 mM ETDA; 1 mM DTT; 0.2 mg/ml BSA; 50% glycerol. |
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| Specificity: |
Mouse Aag catalyzes the excision of the following forms of DNA damage: 3-methyladenine, 3-methylguanine, 7-methylguanine, hypoxanthine, 1,N 6 ethanoadenine, and deoxyinosine. It may also function on the following forms of DNA damage: 7- and 3-ethylpurines, 1-car-boxyethyladenine, 7-carboxyethylguanine, O 2 -methylpyrimidines, 7(2-ethoxyethyl)guanine, 7(2-hydroxyethyl)guanine, 7(2-chloroethyl)guanine, 1,2-bis(7-guanyl)ethane, 3-ethylth-ioethylpurines, N2,3-ethenoguanine, N2,3-ethanoguanine, 5-hydroxymethyluracil, 5-formyl-uracil, 3,N 4 -ethenocytosine, 1,N 2 -ethenoguanine, 3,N 2 -ethenoguanine, chloroacetaldehyde cyclic adducts.
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