| Source: |
Purified from E. coli containing a recombinant plasmid harboring the human FEN-1 gene. |
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| Unit Definition: |
One unit is the amount of enzyme required to cleave a flap DNA structure in a DNA duplex at the rate of 1 pmol/hour at 37°C.
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| Assay Conditions: |
1X REC™ Reaction Buffer 12 (50 mM
Tris-HCl (pH 8.0), 10 mM MnCl2 , and 1 mM DTT), 1X BSA Additive (0.1
mg/ml BSA, 5% glycerol), 50 µl of the above 32P Flap substrate,
and serial dilutions of enzyme in a reaction volume of 10 µl,
incubated for 1 hour at 30°C. The reactions were stopped by the
addition of 5X REC™ Loading Buffer (Cat# 4018-250) and products
are resolved by 20% denaturing polyacrylamide gel electrophoresis.
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| Applications: |
Excision repair research Double-strand break research
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| Storage: |
Freeze in working aliquots at -20°C in a manual defrost freezer to avoid repeated freeze-thawing. The enzyme is supplied in buffer containing 50% glycerol. |
