Fig 4. Detection of BaMV and Ac by dot blot analysis using GfED-tagged antibodies.

(a) Schematic diagram (not drawn to scale) of the proteins involved in panel b. (b) BaMV particles with 5-fold serial dilution (starting at 1000 μg/ml) were loaded (100 μl/well) on nitrocellulose (NC) membrane and overlaid with GfED, then various amounts of anti-BaMV CP IgG (20, 40, 60, 80, and 100 μg) were added and incubated for 2 hr. The results were observed under 450 nm blue light excitation with an orange filter. BaMV, purified viral particles of BaMV; TBS, NC membrane overlaid with TBS buffer as a negative control. (c) Schematic diagram of the proteins involved in panel d. (d) Bacterial samples with 5-fold serial dilution (starting at 10⁹ CFU/ml) were loaded (100 μl/well) on NC membrane and overlaid (incubated) with 10 ml GfED tagged anti-Ac or mAb-29 (80 μg). The results were examined under the illumination of blue light (450 nm excitation), or daylight, and verified by using traditional dot-blotting with anti-Ac and alkaline phosphatase-conjugated anti-rabbit IgG (anti-Rb AP) antibody, followed by color development with NBT/BCIP. Infected, crude extracts from muskmelon leaves infected with Ac (0.1 g leaf tissues in 1 ml TBS as the starting material); Aac31, A. citrulli Aac31; OAC1, A. cattleyae OAC1.