General description
Endonuclease VIII (UniProt: P50465; also known as DNA glycosylase/AP lyase Nei, DNA-(apurinic or apyrimidinic site) lyase Nei, EC:3.2.2.-, EC:4.2.99.18) is encoded by the nei gene (Gene ID: 945320) in E.coli. Endonuclease VIII is a DNA repair enzyme that plays a significant role in maintaining genomic integrity by excising oxidized pyrimidines, such as thymine glycol (Tg) and 5,6,dihydrothymine (DHT) from DNA. The mechanism used by Endo VIII to cleave the DNA backbone (lyase activity) is a β- elimination, resembling the action of form-amidopyrimidine-DNA glycosylase (fpg) protein. It comprises C-terminal FPG-type zinc finger (aa 229-263). It catalyzes hydrolysis of the N-glycosidic bond and β, -elimination of 3 - and 5 -phosphate groups of an apurinic/apyrimidinic site. Nei exhibits a preference for substrates such as thymine glycol and 5,6-dihydrouracil, indicating its specialized role in base excision repair, as evidenced by increased sensitivity of nei mutants to DNA-damaging agents. 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. Single mutants of Endo VIII has been analyzed to elucidate the role of some amino acid residues of Endo VIII in the recognition of DNA lesions and, it has been suggested that, Leu70 plays the role of a DNA lesion sensor , which is used by Endo VIII for recognition of a DNA damage site. This ZooMAb® recombinant monoclonal antibody, generated by our proprietary technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Kladova, OA., et al. (2017). Genes. 8(5);140; Kuznetsov, NA., et al. (2012). DNA Repair. 11(11);884-91).
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
His-tagged, full-length recombinant Endonuclease VIII from Escherichia coli (strain K12).
Application
Quality Control Testing
Evaluated by Western Blotting with full length, recombinant Endonuclease VIII.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected full-length recombinant Endonuclease VIII.
Tested Applications
Affinity Binding Assay: A representative lot of this antibody bound Endonuclease VIII recombinant protein with at least one hundred thousand-fold (100,000X) higher affinity than with non-specific GST recombinant protein.
Enzyme Immunoassay: A representative lot of this antibody detected Endonuclease VIII protein in ELISA application.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 2E5 is a ZooMAb® recombinant rabbit monoclonal antibody that specifically detects Endonuclease VIII.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Other Notes
We are committed to bringing you greener alternative products, which adhere to one or more of the 12 Principles of Green Chemistry. This antibody is preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed, and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency".
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Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
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.