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Index / Trevigen / E. coli Fpg FLARE Module / Product Detail : 4040-100-FM E. coli Fpg FLARE Module
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Dec
3th

#4040-100-FM E. coli Fpg FLARE Module

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  Price : 392   EUR
445   USD
304   GBP
1649   Zloty
52532   JPY
3027   NOK
3244   SEK
444   CHF

Product name : E. coli Fpg FLARE Module

Catalog number : 4040-100-FM

Quantity: >100 Samples

Availability: Yes

Supplier name : Trevigen

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More Details about

Description: Fpg releases damaged bases preferentially from duplex DNA. It has an associated class I AP lyase activity, leaving both 3' and 5' phosphoryl groups. This results from a b, d elimination reaction at the AP sites, producing a single nucleotide gap in the DNA. The enzyme consists of 269 amino acids with a molecular weight of 30.2 kDa.
Source: Purified from E. coli containing a recombinant plasmid harboring the E. coli fpg gene.
Unit Definition: One Unit cleaves 1 pmole of a labeled oligonucleotide probe containing 8-oxoguanine, within an oligonucleotide duplex in one hour at 37°C.
Specificity: Fpg catalyzes the excision of the following forms of DNA damage:
1. Open ring forms of 7-methylguanine, including 2,6-diamino-4-hydroxy-
5-N-methylformamidopyrimidine and 4,6-diamino-5-amidopyrimidine, a lethal lesion.
2. 8-oxoguanine, a highly mutagenic lesion and probably the most important biological substrate of Fpg.
3. 5-hydroxycytosine
4. 5-hydroxyuracil
5. Aflatoxin-bound imidazole-ring-opened guanine
6. Imidazole ring opened N-2-aminofluorene-C8-guanine
Assay Conditions & Analysis: 1X REC Buffer 10 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 10 mM EDTA, and 0.1 mg/ml BSA), 4 pmoles of labeled 8-oxo-dG oligonucleotide annealed to the compliment oligonucleotide, and serial dilutions of enzyme in a reaction volume of 20 μl are incubated for 1 hour at 37°C. For analysis, 20 μl of 2X Loading Buffer (20 mM EDTA, 95% formamide, and 0.13% bromophenol blue) are added, the samples are heated to 95°C for 10 min then fast cooled to 4°C, and the cleavage products are resolved by 20% denaturing polyacrylamide gel electrophoresis, and percent cleavage quantified.
Storage Buffer: 20 mM Tris-Cl (pH 7.8), 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 50% (v/v) glycerol.
Storage Conditions: Store at -20°C in a manual defrost freezer. For long term storage, freeze in working aliquots at -80°C. Avoid repeated freeze-thawings. May be diluted in 10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 1 mM EDTA, 1 mM DTT, 0.1 mg/ml BSA, 50% glycerol and store at -20°C for up to 1 week. Otherwise, dilute enzyme in 1X REC Buffer 10 and use immediately. It is stable for up to 8 hours at 37°C without any loss in activity.

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