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. 2017 Feb 15;114(10):2580–2585. doi: 10.1073/pnas.1614302114

Fig. S3.

Fig. S3.

Purification of WT and E60AE61AE64A HuLf proteins. (A) Anionic exchange chromatograms obtained by a Q-Sepharose Fast Flow column with a 0 to 1 M NaCl gradient (dashed green line). In red and cyan, the elution profiles of WT and E60AE61AE64A HuLf, respectively, followed by UV detector (FPLC; Akta system). (B) SDS/PAGE gel of fractions obtained by anionic exchange chromatography. Lane M, marker; lanes 1 to 6, main peak of WT HuLf profile; lanes 8 to 13, main peak of E60AE61AE64A profile (bands around 20 kDa corresponding to the protein monomers). (C) Size exclusion elution chromatograms (UV measure) of WT and E60AE61AE64A HuLf superimposed with one of an already-characterized ferritin (violet curve; HuHf sample) to demonstrate their 24-meric conformation. (D) SDS/PAGE analysis of fractions eluted with HiLoad 16/60 Superdex 200 column. Lanes M, marker; lanes 1 to 6 main peak of WT HuLf profile; lanes 7 to 12, main peak of E60AE61AE64A profile (bands around 20 kDa corresponding to the protein monomers).