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Endonuclease VIII
| No | Product Name | Cat.# | Conc | Size | Price(￦) | Add |
|---|---|---|---|---|---|---|
| 1 | Endonuclease VIII | M034S | 10 unit/μl | 1,000 units | ￦ 81,000 |
|
| 2 | Endonuclease VIII | M034L | 10 unit/μl | 5,000 units | ￦ 324,000 |
|
Product
description
Endonuclease VIII from E. coli acts as both an N-glycosylase and an AP-lyase. The
N-glycosylase activity releases damaged pyrimidines from double-stranded DNA,
generating an apurinic (AP site). The AP-lyase activity cleaves 3’ and 5’ to
the AP site leaving a 5’ phosphate and a 3’ phosphate. Damaged bases recognized
and removed by Endonuclease VIII include urea, 5, 6- dihydroxythymine, thymine
glycol, 5-hydroxy-5- methylhydanton, uracil glycol, 6-hydroxy-5,
6-dihydrothymine and methyltartronylurea. While Endonuclease VIII is similar to
Endonuclease III, Endonuclease VIII has β and δ lyase activity while
Endonuclease III has β lyase activity.
Characteristics
- Isolated from a recombinant source
- Reaction temperature: 37℃
- Heat inactivation: 75℃ for 10 min
Applications
- Studies of DNA damage and repair
- Single cell electrophoresis (comet assay)
- DNA structure research
- Alkaline unwinding
Quality
control
- Purity: >99% on SDS-PAGE
- Endonuclease-free
- Exonuclease-free
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℃ in 1X Endonuclease VIII 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 min at 37℃.)
Storage
buffer
10 mM Tris-HCl (pH 8.0), 250 mM NaCl, 0.1 mM
EDTA, 50% Glycerol
Reaction
buffer (10X)
100 mM Tris-HCl (pH 8.0), 750 mM NaCl, 10 mM EDTA
Standard reaction conditions
1X Endonuclease VIII Reaction Buffer, Incubate at 37℃
Figure 1. Cleavage of DNA containing an AP site by Endonuclease VIII
20 pmol of 5’ end-labeled DNA substrates were incubated with Endonuclease VIII for 1 hr at 37℃. Reaction products were analyzed on 15% denaturing PAGE.
