| Literature DB >> 21870259 |
Jean-Christophe Amé1, Thomas Kalisch, Françoise Dantzer, Valérie Schreiber.
Abstract
The purification of Poly(ADP-ribose) polymerases from overexpressing cells (Sf9 insect cells, Escherichia coli) has been updated to a fast and reproducible three chromatographic steps protocol. After cell lysis, proteins from the crude extract are separated on a Heparine Sepharose™ column. The PARP-containing fractions are then affinity purified on a 3-aminobenzamide Sepharose™ chromatographic step. The last contaminants and the 3-methoxybenzamide used to elute the PARP from the previous affinity column are removed on the high-performance strong cations exchanger Source™ 15S matrix. The columns connected to an ÄKTA™ purifier system allow the purification of PARPs in 3 days with a high-yield recovery. As described in the protocol, more than 11 mg of pure and highly active mouse PARP-2 can be obtained from 1 L of Sf9 insect cell culture.Entities:
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Year: 2011 PMID： 21870259 DOI： 10.1007/978-1-61779-270-0_9
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745