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The cross-reaction of E. coli ACP with Z and Bacillus ACP inhibited the biosynthesis of ACP-dependent fatty acids. Immunoaffinity chromatography using the anti-E. coli ACP bound to Sepharose resulted in extensive single-step purification of crude preparations of ACP from E. coli and from E. gracilis strain Z and variety bacillaris. The purified ACP was biologically active.