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Fig. 2. Purification of His6-CAP1. HEK293 cells were transfected with pUSH-CAP1. The cell lysate was prepared (lane 2), and His6-CAP1 was successively enriched upon a Ni2+-NTA- (lane 3) and a SP-Sepharose column (lane 4). Although gel filtration through a Superdex 200 column could not separate bound G-actin from His6-CAP1 (lane 5), bound actin was liberated by treating the Ni2+-column-bound His6-CAP1-actin complex with urea. Pure His6-CAP1 was then eluted with imidazole after the urea was removed (lane 6). Molecular size standards were run on lane 1.





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