| Source: |
Purified from E. coli containing a recombinant plasmid harboring the E. coli nfo gene. |
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| Unit Definition: |
One unit is the amount of enzyme required to cleave an AP-site oligonucleotide within an oligonucleotide duplex at the rate of 1 pmol/hour at 37°C. |
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| Assay Conditions: |
1X RECTM Buffer 1 (10 mM HEPES-KOH, pH 7.4, 100 mM KCl), 4 pmoles AP-site oligonucleotide (Cat# 3851-100-01) labeled with 32P, 4 pmoles complementary oligonucleotide (Cat# 3851-100-02), and 0.01-1 Unit (serially diluted) endonuclease IV in a 20 µl reaction volume and incubate for 1 hour at 37°C. Cleavage products are resolved by 20% denaturing PAGE. |
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| Specificity: |
Endonuclease IV exhibits AP endonuclease and 3’- repair diesterase activities. |
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| Applications: |
CometAssayTM/FLARETM
Supercoiled DNA relaxation assay
Alkaline elution assay
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| Storage: |
Freeze in working aliquots at -80°C to avoid repeated freeze-thawing. The enzyme is supplied in 10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 0.1 mg/ml BSA, and 50% glycerol. |
