Fig. 1. Covalent coupling of FITC onto the capsid of AAV2 via primary amino groups. (A) 1012 vg of AAV2-GFP vectors were added to a solution of FITC (3 × 105 eq.) in TBS buffer (pH 9.3) and incubated for 4 h at RT. (B) The procedure was carried out with fluorescein (3 × 105 eq.) as a control. (C–E) 109 vg of each condition was analyzed by dot blot using the A20 antibody that recognizes the assembled capsid (C) or using an anti-FITC antibody (D) or by direct fluorescence emission (E). (F and G) 5 × 108 vg of the same samples were analyzed by western blot using a polyclonal antibody to detect denatured AAV capsid proteins (F) or using an anti-FITC antibody (G). (H) 1010 vg of each condition was analyzed by silver nitrate staining.
