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The 14-3-3σ protein (full-length for fluorescence anisotropy assays, ΔC for crystallography) with an N-terminal His6 tag was expressed in BL21(DE3)-competent Escherichia coli (Novagen) from a pPROEX HTb expression vector. The 14-3-3σ protein (full-length for SPR) with an N-terminal His6-tag and C-terminal Twinstrep-tag was expressed in BL21(DE3)-competent E. coli (Novagen) from a pET28a expression vector.
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