The purification of poly(ADP-ribose) polymerase-3 (PARP-3) from overexpressing cells (Sf9 insect cells, Escherichia coli) has been updated to a fast and reproducible two chromatographic steps protocol. After cell lysis, PARP-3 protein from the crude extract is affinity purified on a 3-aminobenzamide Sepharose™ chromatographic step. The last contaminants and the 3-methoxybenzamide used to elute PARP-3 from the previous affinity column are removed on the high-performance strong cations exchanger MonoQ™ matrix. This step allows also the concentration of the protein. The columns connected to an ÅKTA™ purifier system allow the purification of the protein in 3 days with a high-yield recovery. As described in the protocol, more than 3 mg of pure and active human PARP-3 can be obtained from 1.5 L of E. coli culture.
CITATION STYLE
Amé, J. C., Camuzeaux, B., Dantzer, F., & Schreiber, V. (2017). Purification of recombinant human PARP-3. In Methods in Molecular Biology (Vol. 1608, pp. 373–394). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6993-7_24
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