Brand:  VPCIR Enzymes

SKU: 10010001 Category: Tags: , GTIN: 5744002620006

Human TOP1 Enzyme

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350.00 + excl. VAT

Human topoisomerase 1 (hTOP1) is a 91 kDa type IB topoisomerase.

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

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.

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Certificate of Analysis for Human topoisomerase 1 (hTOP1).

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