APE1

Description

Description:
APE1, also known as HAP1 or Ref1, acts as an AP lyase by hydrolyzing the phosphodiester backbone at the 5′-end of an apurinic (AP) site, generating a 1 base gap in the DNA duplex and leaving 3′-hydroxyl and 5′-deoxyribose phosphate termini. Evidence suggests that APE1 may exhibit weak DNA 3′-diesterase, 3′ to 5′-exonuclease and RNase H activities ^(1-4).

Specific Activity: 2,000,000 U/mg

Applications:

• Single cell gel electrophoresis (Comet assay)
• Alkaline elution
• Alkaline unwinding
• Modified nick translation

Source:
An E. coli strain which carries the cloned human APE1 gene

Supplied in:
10 mM Tris-HCl
50 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 8.0 @ 25°C

Supplied with:
10x Green Buffer

10x Green Buffer:
200 mM Tris-Acetate
500 mM Potassium Acetate
10 mM Dithiothreitol
100 mM Magnesium Acetate
pH 7.9 @ 25°C

Unit Definition:
One unit is defined as the amount of enzyme required to cleave 20 pmol of a 34-mer oligonucleotide duplex contain-ing a single AP site in 1 hour at 37°C.

Recommended Storage Condition: -20ºC

Reference:
1. Demple, B. et al. (1991) Proc. Natl. Acad. Sci. USA, 88, 11450-11454.
2. Barzilay, G. et al. (1995) Nucl. Acids Res., 23, 1544-1550.
3. Barzilay, G. et al. (1995) Nature Struc. Biol., 2, 451-468.
4. Unpublished observations (See “Notes” Section)

Additional information

Manual & Protocols

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