Simultaneous Purification of DNA Topoisomerase I and II from Eukaryotic Cells
- 1 January 1989
- journal article
- research article
- Published by Taylor & Francis in Preparative Biochemistry
- Vol. 19 (1) , 37-48
- https://doi.org/10.1080/10826068908544895
Abstract
We have developed a procedure for the simultaneous purification of DNA topoisomerase I and II from calf thymus. Both enzymes were first extracted from isolated nucleoprotein complexes. After batchwise chromatography on hydroxylapatite the two enzyme activities were separated on a FPLC phenylsuperose column. The enzymes were further purified by a second chromatography on phenylsepharose (topo I) or FPLC Mono Q (topo II). The purification can be finished within three days, yielding 0.5-1.0 mg quantities of homogeneous, enzymatic active preparations of the two proteins from 200 g of starting material.Keywords
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