APE 1

Catalog # Concentration Size List Price Quantity Your Price
M0282L 10000 units/ml 5000 units $483.00
$434.70
M0282S 10000 units/ml 1000 units $119.00
$107.10
Catalog # Size List Price Your Price
M0282L 5000 units $483.00
$434.70
M0282S 1000 units $119.00
$107.10
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • Apurinic/apyrimidinic Endonuclease 1
  • DNA AP endonuclease that catalyzes the cleavage of DNA phosphodiester backbone at AP sites via hydrolysis leaving a 1 nucleotide gap with 3'-hydroxyl and 5' deoxyribose phosphate (dRP) termini
  • APE 1 has also been reported to have DNA 3'-diesterase, 3' to 5' exonuclease, and RNase H activities
  • Also known as HAP1 or Ref-1
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Human apurinic/apyrimidinic (AP) endonuclease, APE 1, also known as HAP 1 or Ref-1, shares homology with Escherichia coli exonuclease III protein. APE 1 cleaves the phosphodiester backbone immediately 5´ to an AP site, via hydrolytic mechanism, to generate a single-strand DNA break leaving a 3´-hydroxyl and 5´-deoxyribose phosphate terminus. Besides AP endonuclease activity, APE 1 has also been reported to have weak DNA 3´ -diesterase, 3´ to 5´ exonuclease and RNase H activities (3-5).

In addition to DNA repair activity, APE 1 is also capable of regulating the DNA binding activity of many transcription factors in vitro by a redox mechanism (Ref-1). As part of this process, APE 1 has been shown to stimulate the DNA binding activity of Fos-Jun heterodimers, Jun-Jun homodimers and Hela cell AP-1 proteins as well as that of several other transcription factors including NF-kB, Myb and members of the ATF/CREB family (7-9).


Product Source
An E. coli strain which carries the cloned human APE 1 gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  APE 1 M0282SVIAL -20 1 x 0.1 ml 10,000 units/ml
  NEBuffer™ 4 B7004SVIAL -20 1 x 1.25 ml 10 X
  APE 1 M0282LVIAL -20 1 x 0.5 ml 10,000 units/ml
  NEBuffer™ 4 B7004SVIAL -20 1 x 1.25 ml 10 X
Application Features
  • Single cell gel electrophoresis (Comet assay)
  • Alkaline elution
  • Alkaline unwinding
  • Modified nick translation

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 20 pmol of a 34 mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 µl in 1 hour at 37°C.

*An AP site is created by treating 20 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C.

Reaction Conditions

1X NEBuffer™ 4
Incubate at 37°C

1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM NaCl
1 mM DTT
0.05 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 8 @ 25°C

Heat Inactivation

65°C for 20 minutes

Unit Assay Conditions

1X NEBuffer 4 containing 20 pmol of fluorescently labled oligonucleotide duplex in a total reaction volume of 10 μl.

Materials Sold Separately
Notes
  • Recommended Dilution for the Comet Assay: 1:103. For a protocol please visit: http://cometassay.com
  • The enzyme has 50% activity in NEBuffers 1-3. (NEB #B7001, NEB #B7002, NEB #B7003)
References
  • Robson, C.N. and Hickson, D.I. (1991). Nucl. Acids Res.. 19, 5519-5523.
  • Flaherty, D.M. (2001). Am. J. Respir. Cell. Mol. Biol.. 25, 664-667.
  • Demple, B. et al. (1991). Proc. Natl. Acad. Sci. USA. 88, 11450-11454.
  • Barzilay, G. et al. (1995). Nucl.Acids Res.. 23, 1544-1550.
  • Barzilay, G. et al. (1995). NatureStruc. Biol.. 2, 451-468.
  • Gorman, M.A. et al (1997). EMBOJ.. 16, 6548-6558.
  • Xanthoudakis, S. et al. (1992). EMBO J.. 11, 3323-3335.
  • Walker, L.J. et al. (1993). Mol.Cell. Biol.. 13, 5370-5376.
  • Vidal, A.E. (2001). EMBO J.. 20, 6530-6539.
  • Wilson, D.M. III et al. (1995). J. Biol. Chem.. 270, 16002-16007.
Usage Guidelines
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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