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Automated multi-step purification of histidine-tagged proteins from unclarified cell lysates
A. Sjöberg, E. Ancker, A. Bergh, J. Lundqvist, K. Öberg and N. Forsberg
GE Healthcare Bio-Sciences AB, SE-751 84, Uppsala, Sweden
Sponsored by GE Healthcare
Published in July 2011 2011 (p.15) Sponsored,vendor-submitted protocol
HisTrap™ FF crude is a ready to use column, intended for purification of histidine-tagged proteins from unclarified cell lysates. No centrifugation or filtration of the samples is required. The total purification time is thereby significantly reduced, an important advantage when working with many samples.In this study, ÄKTAxpress™, a chromatography system designed for unattended multi step protein purification, was used for the purification of unclarified samples. HisTrap FF crude was used in the first purification step, followed by up to three additional chromatography steps.A comparison between unclarified sample purified on HisTrap FF crude and clarified sample purified on HisTrap FF is presented. Further, purification of a large volume of unclarified lysate, as well as a large amount of target protein, is shown.
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