Fitc conjugated goat anti human igg

For immunofluorescence analysis, chamber slides (Nalgene Nunc) were seeded with Vero cells (1x10⁴) from stock cultures. Cells were grown until they reached approximately 80% confluency after which cells were infected for 2 h with ZIKV at a multiplicity of infection (m.o.i.) of 0.01 so that cells expressed the antigens. Fixed cells on the slides were incubated for 1 h at 37°C with twofold dilutions of sera beginning at 1:100 dilution from the ZIKV-IgG administered mice for 90 min at 37°C in a humidified chamber. After washing thrice with PBS, the cells were incubated for 60 min at 37°C with a FITC-conjugated goat anti-human IgG (Santa Cruz Biotechnology Inc). The additional nuclear staining with 4′,6-diamidino-2-phenylindole (DAPI) was performed at room temperature for 20 min. After each incubation step, the samples were washed once with PBS. The samples were subsequently mounted onto glass slides using DABCO and were viewed under a confocal microscope (LSM710; Carl Zeiss). The resulting images were analyzed using the Zen software (Carl Zeiss).