Fig. 2.

Purification of recombinant 1E8-4b Fab-EEF/FLAG conjugate from E. coli culture supernatant as analysed by SDS-PAGE under non-reducing conditions and immunoblotting (lanes 1–5) or Coomassie Brilliant Blue staining (lane 6). Immunoreactive protein bands were detected using M2 anti-Flag antibody followed by goat anti-mouse alkaline phosphatase conjugated antibody and developed with Fast Red/AS-MX Naphthol reagents. Lane 1: Culture supernatant; lane 2: Supernatant following 60% (v/v) SAS precipitation; lane 3: Low-molecular-weight-protein markers (with numbers to the left in kDa); lane 4: precipitated 1E8-4b Fab protein following 60% (v/v) SAS precipitation; lane 5: pooled and concentrated M2 anti-Flag affinity gel purified 1E8-4b Fab eluates; lane 6: Coomassie stain of lane 5.