Application Features

• Single cell gel electrophoresis (Comet assay) (3,4,5)
• Alkaline elution (6)
• Alkaline unwinding (7)

Unit Definition

One unit is defined as the amount of enzyme required to cleave 1 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 in 1X Endonuclease III Reaction Buffer containing 10 pmol of fluorescently labeled oligonucleotide duplex.

* An AP site is created by treating 10 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 Endonuclease III (Nth) Reaction Buffer
Incubate at 37°C

1X Endonuclease III (Nth) Reaction Buffer
20 mM Tris-HCl
1 mM EDTA
1 mM DTT
(pH 8 @ 25°C)

Usage Concentration

10,000 - units/ml

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
pH 7.4

Heat Inactivation

65°C for 20 min

Unit Assay Conditions

1X Endonuclease III (Nth) Reaction Buffer containing 10 pmol of fluorescently labeled oligonucleotide duplexes in a total reaction volume of 10 µl

1. Dizdaroglu, M., Laval, J. and Boiteux, S. (1993). Biochemistry. 32, 12105-12111.
2. Hatahet, Z. et al. (1994). J. Biol. Chem.. 269, 18814-18820.
3. Singh, N. et al. (1988). Exp. Cell Res.. 175, 184-191.
4. Collins, A. et al. (1993). Carcinogenesis. 14, 1733-1735.
5. Collins, A. et al. (1996). Environ. Health Perspect . 104, 465-469.
6. Pflaum, M. et al. (1998). Free Rad. Res.. 29, 585-594.
7. Hartwig, A. et al. (1996). Toxicol. Lett. 88, 85-90.

1. What is the activity of Endonuclease III in the NEBuffer 1-4?
2. What is the molecular weight of Endonuclease III?
3. What types of damaged bases are recognized and removed by Endonuclease III?
4. Is Endonuclease III a tagged protein?
5. What substrate is used to test Endonuclease III activity?
6. Will Endonuclease III work in CutSmart buffer?

1. Comet Assay - Modified for Detection of Oxidized Bases Using the Repair Endonucleases Fpg, hOGG1 and Endonuclease III (Nth)

• Properties of DNA Repair Enzymes

• DNA Damage and PreCR

• NEBioCalculator

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.

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]

• M0268S_L_v1

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

• Endonuclease III (Nth)
• Endonuclease III (Nth) Reaction Buffer

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For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

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.