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Index / Trevigen / hoGG1 Enzyme & Buffer / Product Detail : 4130-500-EB hoGG1 Enzyme & Buffer
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Aug
25th

#4130-500-EB hoGG1 Enzyme & Buffer

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  Price : 867   EUR
984   USD
673   GBP
3644   Zloty
116086   JPY
6690   NOK
7169   SEK
981   CHF

Product name : hoGG1 Enzyme & Buffer

Catalog number : 4130-500-EB

Quantity: 500 samples

Availability: Yes

Supplier name : Trevigen

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Description: Reactive oxygen species generated from such things as ionizing radiation, cellular metabolism, and chemical genotoxins cause the DNA adducts 7,8-dihydro-8-oxoguanine (8-oxoG) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FaPy). Human 8-oxoguanine DNA glycosylase (hOGG1) catalyzes the removal of the 8-oxoG and FaPy through cleavage of the DNA phosphodiester bond following Schiff base chemistry. hOGG1 does not recognize the C=O of 8-oxoG as expected, but rather recognizes a proton on N7 of the nucleotide. By mispairing with adenine during replication, 8-oxoG gives rise to G:C to T:A transversions, a frequent somatic mutation in human cancers. In contrast, a FaPy lesion leads to termination of replication and, therefore, is not considered a pre-mutagenic lesion.
Source: Purified from E. coli containing a recombinant plasmid harboring the a-hOGG1 gene (nuclear protein).
Unit Definition: One Unit cleaves 1 pmole of a labeled oligonucleotide probe containing 8-oxoG base paired with C within a duplex oligo.
Specificity: The catalytic activity of hOGG1 is dependent upon the base the 8-oxoG is paired with in the order of C>T>G, A. hOGG1 is also catalytically active when FaPy is paired with C. FaPy is only repaired when base paired to cytosine.
Assay Conditions and Analysis: 1X REC Buffer 6 (20 mM Tris-Cl (pH 8.0), 1 mM EDTA, 1mM DTT, 100 μg/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), 1.0 mM DTT, 1 mM EDTA, 100 mM NaCl, 1 mM DTT and 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 to avoid repeated freeze-thawing.

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