Endonuclease III (Nth)
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Endonuclease III (Nth) protein from E. coli acts as both N-glycosylase and a AP-lyase. The N-glycosylase activity releases damaged pyrimidines from double-stranded DNA, generating a basic (AP site).
| SKU | OPTIONS | Price | Quantity | ||
|---|---|---|---|---|---|
| ENTH-100 | 50 ug (1 mg/ml) | Please inquire | |||
| ENTH -200 | 1 mg (1 mg/ml) | Please inquire | |||
| ENTH -300 | 100 mg (10 mg/ml) | Please inquire | |||
| ENTH -400 | 1 g (10 mg/ml) | Please inquire | |||
| ENTH -OEM | Any size | Please inquire |
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Description
Description:
Endonuclease III (Nth) protein from E. coli acts as both N-glycosylase and a AP-lyase. The N-glycosylase activity releases damaged pyrimidines from double-stranded DNA, generating a basic (AP site). The AP-lyase activity of the enzyme cleaves 3´ to the AP site leaving a 5´ phosphate and a 3´ ring opened sugar. Some of the damaged bases recognized and removed by Endouclease III include urea, 5, 6 dihydroxythymine, thymine glycol, 5-hydroxy-5 methylhydanton, uracil glycol, 6-hydroxy-5, 6-dihdrothimine and methyltartronylurea.
Applications:
• Single cell gel electrophoresis (Comet assay)
• Alkaline elution
• Alkaline unwinding
Source:
expressed in E. coli
Purity:
> 95 % as determined by SDS-PAGE
Storage:
-80 Avoid repeated freeze-thaw cycles.
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:
1X Endonuclease III (Nth) Reaction Buffer
Incubate at 37°C
1X Endonuclease III (Nth) Reaction Buffer
20 mM Tris-HCl
1 mM EDTA
1 mM DTT
(pH 8 @ 25°C)
Usage Concentration:
10,000 units/ml
Storage Temperature:
-20°C
Storage Conditions:
10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4
Heat Inactivation:
65°C for 20 min
Additional information
| OPTIONS | 50 ug (1 mg/ml), 1 mg (1 mg/ml), 100 mg (10 mg/ml), 1 g (10 mg/ml), Any size |
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