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This allows the peptide to fully dissolve and stabilize

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METHOD DETAILS Sample preparation FOXO4 FHD (95-195) was subcloned into the pET His6 Tobacco Etch Virus (TEV) LIC cloning vector (2B-T, obtained from Scott Gradia, Addgene Plasmid #29666) and transformed into BL21 (DE3) cells

Cloudiness in the reconstituted solution suggests aggregation, particularly common with hydrophobic peptides

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Pull-down assay Five micrograms of the bait protein c-MET927-TSI3-His was immobilized to TSE resin by incubating for 30 min on ice and washed with binding buffer PBS (Gibco) supplemented with10 M Tinzaparin and 2 mM CaCl2