商品编号


EG-1007



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


DNA Modifying Enzymes



Size

1,000 Units

商品信息



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 (1,2). While Endonuclease VIII is similar to Endonuclease III, Endonuclease VIII has β and δ lyase activity while Endonuclease III has β lyase activity.




Applications



Single cell gel electrophoresis (Comet assay) (3,4,5)

Alkaline elution (6)

Alkaline unwinding (7)





Source

An
E. coli
strain which carries the cloned Endonuclease VIII
nei
gene



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 ?l in 1 hour at 37°C 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 minutes at 37°C.




Components



Endonuclease VIII:
10,000 U/ml in 10 mM Tris-HCl, pH 8.0 @ 25°C, 250 mM NaCl, 0.1 mM EDTA, 50% glycerol

10X Reaction Buffer
: 100 mM Tris-HCl, pH 8.0 @ 25°C, 750 mM NaCl, 10 mM EDTA





Quality Control

The absence of
endonucleases, exonucleases,
E.coli DNA contamination confirmed by appropriate quality tests.



Storage Condition


-20 °C




References



Dizdaroglu, M., Laval, J. and Boiteux, S. (1993).
Biochemistry
. 32, 12105-12111.

Hatahet, Z. et al. (1994).
J. Biol. Chem.
. 269, 18814-18820.

Singh, N. et al. (1988).
Exp. Cell. Res.
. 175, 184-191.

Collins, A. et al. (1993).
Carcinogenesis
. 14, 1733-1735.

Collins, A. et al. (1996).
Environ. Health Perspect
. 104, 465-469.

Pflaum, M. et al. (1998).
Free Rad. Res.
. 29, 585-594.

Hartwig, A. et al. (1996).
Toxicol. Lett.
. 88, 85-90.

Marks, K. unpublished observation.