Fig. 2.

Cell free glycosylation (CFG). A schematic diagram of cell free glycosylation analysis; B western blot analysis of cell free glycosylation upon gradual increase of cell debris containing glycan donor denoted by the grey triangle. Western blot of 5 µg His-tagged CmeA protein purified by nickel affinity chromatography Protein samples were separated by SDS-PAGE 4–12% bis–tris gel (Invitrogen) with MOPS buffer and transferred to nitrocellulose membrane with an iBlot 2 dry blotting system. The membrane was probed with anti-His (Invitrogen) and anti- Ft-O antigen monoclonal antibody (Abcam) and detected with fluorescently labelled secondary antisera (red-His, green-Ft-O-antigen) on a LI-COR Odyssey scanner