Fiche publication
Date publication
janvier 2011
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DANTZER Françoise
,
Dr SCHREIBER Valérie
Tous les auteurs :
Ame JC, Kalisch T, Dantzer F, Schreiber V
Lien Pubmed
Résumé
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 AKTA 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.
Référence
Methods Mol Biol. 2011;780:135-52.
