Brand:  VPCIR Enzymes

SKU: N/A Category: Tags: , GTIN: 5744002620006

Human TOP1 Enzyme

350.00 + excl. VAT

High purity, catalytically active hTOP1 Enzyme overexpressed in yeast and purified to single band homogeneity.

Uses

Research in Cancer, Research in Chronic diseases, Research in Infectious diseases

Purity

80%

Sizes

1.5 µg ≥ 1,500 units, 3.5 µg ≥ 3,500 units, 10 µg ≥ 10,000 units

Variants

Single-pack, 2-pack, 3-pack

Shipping

Not IATA restricted

Storage

Must be stored at -18°C to -20°C upon arrival

Clear

Human topoisomerase 1 (hTOP1) is supplied in Dilution Buffer at a minimum concentration of 30ng/µl, corresponding to at least 30 U/µl. 1 U of hTOP1 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 hTOP1 band has been estimated by densitometry and quantified by using ImageJ software comparing with a protein standard of known concentration. The hTOP1 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 hTOP1 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 non-tagged hTOP1 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 cooled by 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 hTOP1 activity. The enzyme has a lifetime of 12 months when stored under optimal conditions.

Certificate of Analysis for Human topoisomerase 1 (hTOP1).

 

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Safety Data Sheet for Human TOP1 Enzyme products.

QR-code for Human TOP1 Enzyme SDS

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1. Blanco, L.; Bernad, A.; Lázaro, J.M.; Martín, G.; Garmendia, C.; Salas, M. Highly Efficient DNA Synthesis by the Phage ϕ 29 DNA Polymerase. J. Biol. Chem. 1989, 264, 8935–8940, doi:10.1016/s0021-9258(18)81883-x.

2. Yokouchi, H.; Fukuoka, Y.; Mukoyama, D.; Calugay, R.; Takeyama, H.; Matsunaga, T. Whole-metagenome amplification of a microbial community associated with scleractinian coral by multiple displacement amplification using j 29 polymerase. 2006, 8, 1155–1163, doi:10.1111/j.1462-2920.2006.01005.x.

3. Larsson, C.; Koch, J.; Nygren, A.; Janssen, G.; Raap, A.K.; Landegren, U. In situ genotyping individual DNA molecules by target-primed rolling-circle amplification of padlock probes. 2004, 1, 1–6, doi:10.1038/NMETH723.

4. Salas, M.; Blanco, L.; Lázaro, J.M.; De Vega, M. The bacteriophage φ29 DNA polymerase. IUBMB Life 2008, 60, 82–85, doi:10.1002/iub.19.


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