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

IX Endonuclease III (Nth) Reaction Buffer
Incubate at 37°C

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

Usage Concentration

10,000 - units/ml

Storage Temperature

-20°C

Storage Conditions

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

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

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

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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.