hNEIL1

产品编号：M0335L 品牌：New England Biolabs 原厂货号：M0335L
产品分类：Discontinued已停产
应用分类：

包装：

2,500 units

运保温度:

-20°C

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描述：

Description:
Human NEIL1 acts as both an N-glycosylase and an AP-lyase. The N-glycosylase activity releases oxidized pyrimidines and purines 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 hNEIL1 include thymine glycol, 5,6-dihydrothymine, 5,6-dihydroxyuracil, 5-hydroxycytosine, 5-hydroxyuracil, 5-formyluracil, 8oxo-guanine opposite C, T or G, 4,6-diamino-5-formamidopyrimidine and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (1,2).
Source:
An E. coli strain that carries the cloned human NEIL1 gene.

Reagents Supplied:
Endonuclease VIII Reaction Buffer
Heat Inactivation:
65°C for 20 minutes
Reaction Conditions:
1X Endonuclease VIII Reaction Buffer
Incubate at 37°C.

1X Endonuclease VIII Reaction Buffer:
10 mM Tris-HCl
75 mM NaCl
1 mM EDTA
pH 8.0 @ 25°C

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.

*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.

Molecular Weight: 43,685 Daltons

Diluent Compatibility: Diluent Buffer A
50 mM KCl, 10 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, 200 µg/ml BSA and 50% glycerol (pH 7.4 @ 25°C)

Unit Assay Conditions: 1X Endonuclease VIII Reaction Buffer containing 5 pmol of fluorescently labeled oligonucleotide duplex in a total reaction volume of 10 µl.

Storage Conditions:
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

注意事项：

Compared to the activity on AP-site, hNEIL1 has ≥ 50% activity on thymine glycol and ≥ 25% activity on dhU, dhT, 5hoU and 5hoC. hNEIL1 is active on 8oxoG opposite C,T and G, but is not active on an 8oxoG:A base pair. hNEIL1 is not active on 5-hydroxymethyluracil.