Goat anti mouse igg fitc secondary antibody

To visualize EGFP expression
cells were grown on rounded glass coverslips and fixed with methanol
100% for 2 min, permeabilized with 0.01% TritonX (Sigma-Aldrich) in
PBS for 15 min and blocked with 0.1% BSA for 30 min, both at room
temperature. Coverslips were incubated overnight at 4 °C with
a mouse monoclonal antibody against GFP (Sigma-Aldrich, diluted 5
μg/mL) derived from mice immunized with a synthetic peptide
corresponding to amino acids 132–144 of the GFP from jellyfish Aequorea victoria, followed by a goat antimouse IgG-FITC
secondary antibody (Sigma-Aldrich, diluted 1:200) for 1 h at 37 °C.
Nuclei were visualized with 1 μg/mL of 4′,6-diamidino-2-phenylindole
(DAPI) and examined on a Zeiss Axioskop microscope equipped for fluorescence,
images were captured with a CCD digital camera (AxioCam, Zeiss) and
then transferred to Adobe PhotoShop for printing.

The virus was diluted to 10⁷ PFU/ml in DMEM without serum and irradiated in microtiter plates at a distance of 6 cm with shortwave (254 nm) UV light for 20 min (Helentjaris and Ehrenfeld, 1977 (link)). UV-irradiated virus infectivity was tested with a plaque assay. Secondly, UV-inactivated virus was inoculated into A549 cells for 2 h, and they were fixed with cold absolute methanol and kept at −20 °C for 15 min. Viral antigen was detected by indirect immunofluorescence with mouse polyclonal antibody anti-reovirus and followed by goat anti-mouse IgG-FITC secondary antibody (Sigma, St. Louis, MO, USA). The treated viruses were unable to form plaques and viral antigens could be detected by indirect immunofluorescence, which was considered to be UV-inactivated (Soares et al., 2009 (link)).