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Index / Trevigen / E. coli Endonuclease III Enzyme & Buffer / Product Detail : 4045-01K-EB E. coli Endonuclease III Enzyme & Buffer
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Aug
26th

#4045-01K-EB E. coli Endonuclease III Enzyme & Buffer

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  Price : 333   EUR
378   USD
258   GBP
1402   Zloty
44659   JPY
2574   NOK
2758   SEK
377   CHF

Product name : E. coli Endonuclease III Enzyme & Buffer

Catalog number : 4045-01K-EB

Quantity: 1000 Units

Availability: Yes

Supplier name : Trevigen

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More Details about

Description: Endonuclease III is a DNA glycosylase with an associated AP lyase activity. Endonuclease III releases bases damaged by UV light, ionizing radiation, osmium tetroxide, or acid. It cleaves abasic sites by b-elimination, producing a single nucleotide gap in the DNA, and contains an iron-sulfur group which helps to maintain its three dimensional conformation. The enzyme has a molecular weight of 23.4 kDa and is suitable for use in FLARE™.
Source: Purified from E. coli containing a recombinant plasmid harboring the E. coli nth gene.
Unit Definition: One Unit of enzyme cleaves 1 pmole of an oligonucleotide probe containing an AP site within an oligonucleotide duplex in one hour at 37 oC.
Substrate Specificity: Endonuclease III catalyzes the excision of the following forms of
DNA damage: Cis-trans-thymine glycol, 5,6-dihydrothymine, 5,6-dihydroxydihydrothymine, alloxan, urea, uracil, 5-hydroxy-5-methylhydantoin, methyltartronylurea, 6-hydroxy-5,6-dihydropyrimidines, 5-hydroxycytosine, 5-hydroxyuracil, 5-hydroxy-6-hydrothymine, 5,6-dihydrouracil, glycol, 5-hydroxy-6-hydrouracil, and AP sites.
Assay Conditions & Analysis: Enzyme may be diluted in 10 mM HEPES-KOH (pH 7.4) and 100 mM KCl for immediate use. In 1X REC Buffer 4 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 10 mM EDTA), add 4 pmole of a 32P-labeled oligonucleotide containing an AP site, 4 pmole of a complementary oligonucleotide, and serial dilutions of enzyme in a 20 μl reaction volume; incubate for 1 hour at 37 oC. The cleavage products are resolved by 20% denaturing polyacrylamide gel electrophoresis. Bands are cut out and radioactivity counted to quantify the cleavage products.
Storage Buffer: 10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 0.1 mg/ml BSA, and 50% (v/v) glycerol.
Storage Conditions: Store at –20 oC in a manual defrost freezer. For long-term storage, freeze at –80 oC in working aliquots. Avoid repeated freeze-thawing.

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