indirect ELISA: suitable western blot: 10-20 μg/mL using recombinant endonuclease VIII (Cat No. E0651)
isotype
IgM
shipped in
dry ice
storage temp.
−20°C
target post-translational modification
unmodified
General description
Monoclonal Anti- Endonuclease VIII (mouse IgM isotype) is derived from the E8-122 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from mice . Endonuclease VIII (Endo VIII) from E. coli is a product of the nei gene. It is a DNA N-glycosylase and abasic site lyase (AP1 lyase. It comprises C-terminal Cys4-type β/β-antiparallel zinc finger with an arginine residue (Arg-252).
Specificity
Monoclonal Anti-Endonuclease VIII recognizes endonuclease VIII from E. coli (approx. 30 kDa).
Immunogen
recombinant full length endonuclease VIII from Escherichia coli.
Application
Monoclonal Anti-Endonuclease VIII antibody produced in mouse may be used in enzyme-linked immunosorbent assay (ELISA) and immunoblotting.
Biochem/physiol Actions
Endonuclease VIII (Endo VIII) from E. coli is a DNA repair protein that recognizes and removes modified pyrimidines, such as thymine glycol (Tg) and 5,6,dihydrothymine (DHT), from DNA.. Its modified base substrate specificity overlaps the endonuclease III (endo III, nth) substrate specificity. This cross-reactivity is manifested in E. coli mutants. E. coli nth and nei mutants are either not sensitive or are slightly more sensitive to ionizing radiation and hydrogen peroxides than the wild type. The nei-nth double mutant is hypersensitive to oxidative stress. The mechanism used by Endo VIII to cleave the DNA backbone (lyase activity) is a β-δ elimination, resembling the action of formamidopyrimidine-DNA glycosylase (fpg) protein.
Physical form
Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.
Storage and Stability
For continuous use, store at 2-8 °C for up to one month.For prolonged storage, freeze in working aliquots at −20 °C. Repeated freezing and thawing is not recom-mended. Storage in frost-free freezers is also notrecommended. If slight turbidity occurs upon prolongedstorage, clarify the solution by centrifugation beforeuse. Working dilutions should be discarded if not usedwithin 12 hours.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Endonuclease VIII (Nei) of Escherichia coli is a DNA repair enzyme that excises oxidized pyrimidines from DNA. Nei shares with formamidopyrimidine-DNA glycosylase (Fpg) sequence homology and a similar mechanism of action: the latter involves removal of the damaged base followed
Endonuclease VIII (Nei), which recognizes and repairs oxidized pyrimidines in the base excision repair (BER) pathway, is sparsely distributed among both the prokaryotes and eukaryotes. Recently, we and others identified three homologs of Escherichia coli endonuclease VIII-like (NEIL) proteins in
Endonuclease VIII (Nei) excises oxidatively damaged pyrimidines from DNA and shares structural and functional homology with formamidopyrimidine-DNA glycosylase. Although the structure of Escherichia coli Nei is solved [Zharkov et al. (2002) EMBO J. 21, 789-800], the functions of many of
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