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Index / Trevigen / E. coli MutY Enzyme & Buffer / Product Detail : 4000-500-EB E. coli MutY Enzyme & Buffer
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Dec
10th

#4000-500-EB E. coli MutY Enzyme & Buffer

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  Price : 365   EUR
415   USD
283   GBP
1536   Zloty
48953   JPY
2821   NOK
3023   SEK
413   CHF

Product name : E. coli MutY Enzyme & Buffer

Catalog number : 4000-500-EB

Quantity: 500 Units

Availability: Yes

Supplier name : Trevigen

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Description: E. coli MutY acts together with Fpg to prevent the potentially mutagenic consequences of 8-oxo-dG lesions. 8-oxo-dG lesions escaping repair by Fpg frequently pair with A during DNA replication, producing an 8-oxo-dG:A mispair. MutY removes the A from this base pair to initiate base excision repair. In the absence of MutY, DNA replication after an 8-oxo-dG:A mismatch results in thymine incorporation opposite the adenine in one of the daughter strands, creating a fixed mutation. MutY has an associated AP lyase activity.
Source: Purified from E. coli containing a recombinant plasmid harboring the E. coli MutY gene.
Unit Definition: One unit of enzyme cleaves 1 pmole of an oligonucleotide duplex containing an A/G mismatch in 1 hour at 37 oC. Only the strand with the A is cleaved.
Assay Conditions & Analysis: 4 pmoles of A/G mismatch oligonucleotide set with the A oligo end-labeled, 1X REC™ Buffer 4 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 10 mM EDTA), and serial dilutions of enzyme in a 20 μl reaction volume are incubated for 1 hour at 37 oC. To complete cleavage of abasic site, fresh NaOH is added to final concentration of 166 mM then heated for 15 minutes at 95 oC. For analysis, 24 μl of 2X Loading Buffer (20 mM EDTA, 97% formamide, and 0.2% bromophenol blue) is added, the samples are heated at 95 oC for 5 min then fast cooled to 4 oC, and the cleavage products are resolved by 20% denaturing polyacrylamide gel electrophoresis. The bands are analyzed to quantify the cleavage products.
Storage Buffer: 20 mM Tris pH 7.5, 100 mM NaCl, 1 mM EDTA, 1 mM DTT, and 50% (v/v) glycerol.
Storage Conditions: Store at -20 oC in a manual defrost freezer. For long term storage, freeze in working aliquots at -80 oC. Avoid repeated freeze-thawings. Enzyme may be diluted in storage buffer containing 0.1 mg/ml BSA and stored at -20 oC for 2 weeks of experimental use.

 

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