Product Class: Other

APE 1
NEB4 cloned at NEB recombinant 37 65 Heat

Product Introduction

  • 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
Catalog # Size Concentration
M0282S 1000.0 units 10000 units/ml
M0282L 5000.0 units 10000 units/ml

Product Information

Description

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.
This product is related to the following categories:
DNA Repair Enzymes and Structure-specific Endonucleases Products
This product can be used in the following applications:
Polymerases for DNA Manipulation

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

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.

Application Features

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

Product Notes

  1. Recommended Dilution for the Comet Assay: 1:103. For a protocol please visit: //cometassay.com
  2. The enzyme has 50% activity in NEBuffers 1-3. (NEB #B7001, NEB #B7002, NEB #B7003)

References

  1. Robson, C.N. and Hickson, D.I. (1991). Nucl. Acids Res.. 19, 5519-5523.
  2. Flaherty, D.M. (2001). Am. J. Respir. Cell. Mol. Biol.. 25, 664-667.
  3. Demple, B. et al. (1991). Proc. Natl. Acad. Sci. USA. 88, 11450-11454.
  4. Barzilay, G. et al. (1995). Nucl.Acids Res.. 23, 1544-1550.
  5. Barzilay, G. et al. (1995). NatureStruc. Biol.. 2, 451-468.
  6. Gorman, M.A. et al (1997). EMBOJ.. 16, 6548-6558.
  7. Xanthoudakis, S. et al. (1992). EMBO J.. 11, 3323-3335.
  8. Walker, L.J. et al. (1993). Mol.Cell. Biol.. 13, 5370-5376.
  9. Vidal, A.E. (2001). EMBO J.. 20, 6530-6539.
  10. Wilson, D.M. III et al. (1995). J. Biol. Chem.. 270, 16002-16007.

Protocols, Manuals & Usage

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the activity of APE 1 in NEBuffers 1-4?
  2. What is the molecular weight of APE 1?
  3. Is APE 1 a tagged protein?
  4. What substrate is used to test APE 1 activity?

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.