Monkeypox Virus TOP1 Enzyme
350.00 €+ excl. VAT
High purity, catalytically active monkeypox virus TOP1 Enzyme overexpressed in yeast and purified to single band homogeneity.
Research in Cancer, Research in Chronic diseases, Research in Infectious diseases
1.5 µg ≥ 1,500 units, 3.5 µg ≥ 3,500 units, 10 µg ≥ 10,000 units
Single-pack, 2-pack, 3-pack
Not IATA Restricted
Must be stored at -18°C to -20°C upon arrival
Monkeypox Virus TOP1 Enzyme (mpvTOP1) is supplied in Dilution Buffer at a minimum concentration of 30ng/µl, corresponding to at least 30 U/µl. 1 U of mpvTOP1 relaxes 0.5 µg of supercoiled pUC19 when incubated in Reaction Buffer and 150 mM NaCl in a total reaction volume of 20 µl at 37 °C for 30 minutes. The purity and the concentration of the enzyme has been determined by SDS-PAGE followed by Coomassie brilliant-blue staining. The intensity of the mpvTOP1 band has been estimated by densitometry and quantified by using ImageJ software comparing with a protein standard of known concentration. The mpvTOP1 enzyme is free from nucleases contaminations and 80% pure as determined by SDS-PAGE analysis. The enzyme preparation is highly stable and recommended for activity assays, including cleavage-ligation experiments, equilibrium, nicking and relaxation assays. The mpvTOP1 enzyme is supplied together with: • Dilution Buffer: 5 mM Tris-HCl pH 7.5, 400 mM NaCl, 1mM DTT, 50% glycerol. • Reaction Buffer 10x: 100 mM Tris-HCl pH 7.5; 50 mM MgCl2, 50 mM CaCl2; 50 µg/ml BSA. • Nucleases-free water. • 1M NaCl.
Recombinant 6HIS-tagged mpvTOP1 is prepared by overexpressing the enzyme in Saccharomyces cerevisiae followed by purification by affinity chromatography adapted from Knudsen et al, J. Chrom. 1996, DOI: 10.1016/0378-4347(96)00152-1
The enzyme is shipped packed with ice gel packs and must be stored at -18°C to -20°C upon arrival. The temperature must be controlled to avoid that the enzyme preparation solidifies. This would destroy the enzyme integrity and cause a drop of the mpvTOP1 activity. The enzyme has a lifetime of 12 months when stored under optimal conditions.
Certificate of Analysis for Monkeypox Virus Topoisomerase 1 (mpvTOP1).
Safety Data Sheet for Monkeypox Virus TOP1 Enzyme products.
- Estep, R.D.; Messaoudi, I.; O’Connor, M.A.; Li, H.; Sprague, J.; Barron, A.; Engelmann, F.; Yen, B.; Powers, M.F.; Jones, J.M.; et al. Deletion of the Monkeypox Virus Inhibitor of Complement Enzymes Locus Impacts the Adaptive Immune Response to Monkeypox Virus in a Nonhuman Primate Model of Infection. J. Virol. 2011, 85, 9527–9542, doi:10.1128/jvi.00199-11.
- Shuman, S. Vaccinia virus DNA topoisomerase: A model eukaryotic type IB enzyme. Biochim. Biophys. Acta – Gene Struct. Expr. 1998, 1400, 321–337, doi:10.1016/S0167-4781(98)00144-4.
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