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Fig. 5 | Microbial Cell Factories

Fig. 5

From: Screening and purification of nanobodies from E. coli culture supernatants using the hemolysin secretion system

Fig. 5

Disulphide bond formation in Nb-HlyA fusions. Thiol alkylation assay to determine the oxidation state of Cysteines (Cys) in secreted and cytoplasmic Nb-HlyA fusions from TD4 and TF1 clones, containing 2 and 4 Cys residues, respectively. Secreted proteins in culture supernatants and bacterial samples are treated (+) or not (−) with AMS (alkylating agent) and DTT (reducing agent), as indicated. Retarded mobility of alkylated polypeptides is indicative of reduced thiol groups. Protein bands of Nb-HlyA fusions developed after non-reducing SDS-PAGE and Western blot with anti-E-tag mAb

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