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Cat. No.: ENN8-1k (for 1KU)

Cat. No.: ENN8-5k (for 5KU)

Cat. No.: ENN8-20k (for 20KU)

Description

Endonuclease VIII is a DNA repair enzyme with N-glycosylase activity and AP-lyase activity. The N-glycosylase activity of Endonuclease VIII cleaves and releases damaged pyrimidine bases from the double-stranded DNA, creating an apurinic (AP) site. The AP-lyase activity of Endonuclease VIII can cleave the 3' and 5' ends of the AP site, removing the AP site and generating a gapped DNA with 3' and 5' phosphate ends.

The damaged bases that can be recognized and cleaved by Endonuclease VIII include urea, 5,6-dihydroxythymine, thymine glycol, 5-hydroxy-5-methylhydantoin, uracil glycol, 6-hydroxy-5,6-dihydrothymine, and methyltartronylurea [1,2]. Although Endonuclease VIII is similar to Endonuclease III, it possesses both β and δ-lyase activities, while Endonuclease III only has β-lyase activity.

Applications

• Used for DNA damage and repair research.
• Single-cell gel electrophoresis (comet assay).
• Alkaline elution.
• Alkaline denaturation.
• Recognition and cleavage of damaged pyrimidine bases on double-stranded DNA.
• Cleavage of the 3' and 5' ends of AP sites, resulting in a gapped DNA with 3' and 5' phosphate ends.

Source

Obtained through expression and purification in Escherichia coli.

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 within 1 hour at 37°C in 1X Endonuclease VIII Reaction Buffer containing 10 pmol of fluorescently labeled oligonucleotide duplex.

Purity

Greater than 95%, devoid of DNA endonuclease and other exonuclease activities, and lacks RNAse activity.

Enzyme Storage Solution

10 mM Tris-HCl (pH 8 @ 25°C), 250 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% (v/v) Glycerol.

Inactivation or Inhibition

Heating at 75°C for 10 minutes renders Endonuclease VIII inactive.

10X Endonuclease VIII Buffer

100 mM Tris-HCl, 750 mM NaCl, 10 mM EDTA, pH 8 @ 25°C.

Precautions

• When using this product, it should be stored in an icebox or on ice, and after use, it should be immediately stored at -20°C.
• This product is intended for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in residential households.
• For your safety and health, please wear laboratory attire and disposable gloves when handling.

Storage

The minimum shelf life is 2 years at -20°C.