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Human Topoisomerase II, Alpha from USB Corp.

ProductsHuman Topoisomerase II, Alpha from USB Corp.
Company USB Corp.
Item Human Topoisomerase II, Alpha
Price 
Description Source: E. coli containing a clone of the human Topoisomerase II gene.
Description:Topoisomerase II alters the topological state of nucleic acids by passing an intact DNA helix through a transient break which generates a separate DNA helix(1,2). As a result of its double-stranded DNA passage mechanism, the enzyme can relax negatively or positively supercoiled DNA, as well as catenate/decatenate or knot/unknot DNA molecules. Topoisomerase II has an absolute requirement for divalent cation and ATP (or dATP).
Tested User Friendly
Properties:
Molecular Weight: 340,000 Da homodimer
Optimum Temperature: 3037C
Optimum pH: 7.9, rapidly loses activity below pH 7.5
Requirements for Divalent Cation: 5mM Mg2+ is optimum for catalytic activity and 5mM Ca2+
 for DNA cleavage.
Optimum ATP Concentration: 0.5 to 1.0mM
Optimum Ion Strength: 100-170mM KCl/NaCl
Inhibitors: N-ethylmaleimide, zinc, novobicin, coumermycin A1, etoposide, amsacrine
Purity:Greater than 98% pure as determined by silver-stained SDS-PAGE. Free of contaminating exonucleases and endonucleases.
Storage Buffer:15mM sodium phosphate (pH 7.1), 700mM NaCl, 0.1mM EDTA, 0.5mM DTT, 50% glycerol.
Shipping and Storage:
Shipped on dry ice. Store at -20C for frequent use. Store at -80C for long-term storage.
Assay Conditions:The reaction mixture contains 10mM Tris-HCl, pH 7.9, 175mM KCl, 0.1mM EDTA, 5mM MgCl2, 2.5% glycerol, 0.5mM ATP.
Unit Definition:One unit is the amount of enzyme required to fully relax 0.3 g (5nM) of negatively supercoiled pBR322 plasmid DNA in 30 minutes at 37C under the standard assay conditions.
Concentration:20 units/l
Tested User FriendlyTM Functional Test: Tested for activity in standard Topoisomerase II unit assay.
Applications:*Relaxing negatively or positively supercoiled DNA.
*Catenating or decatenating DNA.
*Knotting or unknotting DNA.
PROTOCOL FOR RELAXATION OF SUPERCOILED DNA:
Topoisomerase II, Alpha and 10X Topoisomerase II Reaction Buffer (PN 73592) are functionally tested in the following protocol using 300 ng (0.1 pmol) pBR322 DNA at 30C.
*Mix
10X Topo II Reaction Buffer 2 l
Supercoiled DNA 0.11.0 pmol
Topoisomerase II, Alpha 110 units
H2O to 20 l
*Mix and incubate at 3037C for 15 minutes.
*Stop the reaction by the addition of 3 l of 7mM EDTA or by the addition of SDS
to 1%.
Info USB Corp.USB Corp.
26111 Miles Road
Cleveland Ohio 44128


Call USB Corp. to buy products (US and Canada only; other locations use numbers below)
USA Canada 1-216-245-1829
Customer Service: 216.765.5000
Fax Number: 216.464.5075
Web Site: http://www.usbweb.com
NOTE: Price information is approximate list price and actual prices may vary.

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