FIGURE 2.

Antibody biotinylation. A, SEC-HPLC elugram of IMAC purified full-length bisbiotinylated chimeric antibody 833cb. B, Western analysis by SA-HRP conjugate of 833cb versus polyclonal human IgG; only the heavy chain is biotinylated as seen in reducing (R) conditions (NR, non-reducing conditions). C, expression of BirA in 293 cells (in green, left panel) follows a perinuclear membranous network that extends deep in the cytoplasm and substantially colocalizes with the Golgi network (in red, all panels); most of antibody-overexpressing 293 cells have an enlarged Golgi network staining that colocalizes with expression of 833cb (in green, right panel).