Figure 4: Site-directed mutagenesis and purification. | Scientific Reports

Figure 4: Site-directed mutagenesis and purification.

From: A hydrophobic patch surrounding Trp154 in human neuroserpin controls the helix F dynamics with implications in inhibition and aggregation

Figure 4

0.8% agarose gel image of PCR amplification of (a) W154A and (b) W154P using plasmid containing WT NS gene as a template. 12% SDS-PAGE image of Ni-NTA purification of (c) WT NS, (d) W154A and (e) W154P Lane M, 2, 3, 4, 5 and 6 corresponds to protein marker, washing with high salt buffer, washing with 50 ml buffer A, washing with 20 mM Tris-HCl [pH 8.0], elution with 300 mM imidazole in buffer A and elution with 600 mM imidazole in buffer A, respectively. Fractions of Lane 5 and 6 were pooled, buffer exchanged and concentrated to run on Gel-filtration column to remove impurities.

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