| Source: |
Purified from E. coli containing a recombinant plasmid harboring the E. coli xth A gene. |
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| Unit Definition: |
One unit is the amount of enzyme required to produce 1 nmole of acid-soluble total nucleotide in 30 minutes at 37°C. |
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| Assay Conditions: |
1X RECTM Buffer 1 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl), 0.5 pmol AP-oligonucleotide (Cat# 3851-100-01) labeled with 32P, 0.5 pmole complementary oligonucleotide (Cat#3851-100-02), and 5 units of Exonuclease III in a 20 µl reaction volume. Following incubation for 1 hour at 37°C, cleavage products are resolved by 20% denaturing polyacrylamide gel electrophoresis. |
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| Specificity: |
E. coli exonuclease III cleaves 5’ to AP sites. It also has 3’ diesterase, 3’- to 5’- exonuclease, and RNase H activities. The enzyme is stimulated by Mg2+ and requires double-stranded
substrates. |
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| Applications: |
CometAssayTM/FLARETM
Supercoiled DNA relaxation assay
Alkaline elution assay
|
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| Storage: |
Freeze at -80°C in working aliquots to avoid repeated freeze-thaw. The enzyme is supplied in a buffer containing 50% glycerol. |
