Fig 2. DomI purification by cation exchange chromatography.

(A) The dialyzed sample was loaded into a HiPrep CM FF column, previously equilibrated with sodium acetate buffer (20 mM, pH 5.0), at 3 mL/min flow rate. Elution was performed by a jump in ionic strength (acetate buffer 20 mM, pH 5.0, NaCl 1M). Absorbance at 280 nm (black solid trace) and conductivity (red dashed trace) were measured. (B) SDS-PAGE analysis. The flowthrough and the bound fraction eluted from the column were concentrated using ultrafiltration devices before loading. Empty lanes were cropped. (C) UV-Vis spectra of the different fractions, dialyzed sample (blue dashed trace), flowthrough (green dotted trace) and bound fraction eluted from the column (black solid trace). Dilutions with similar 280 nm absorbance were prepared.