Gold conjugated goat anti rabbit igg

PIE monolayers were first incubated with SADS-CoV at an MOI = 200 for 30 min at 4°C, then transferred to 37°C, fixed with immune electron microscopy fixative (0.1% glutaraldehyde and 3%PFA in 0.1 M PBS) for 2 h at room temperature, and finally collected. The collected cell precipitate was resuspended and washed twice with precooled 0.1 M PBS (pH 7.4). After the supernatant was removed, samples were processed with dehydration, resin penetration, embedding and polymerization steps. Samples were sliced into 90-nm ultrathin cryosections using a Leica UC7 ultramicrotome and collected onto 150-mesh nickel grids for immunogold labeling. The nickel grids were incubated with a 1:20 dilution of rabbit anti-caveolin-1 antibody (A1555, ABclonal) overnight at 4°C. The nickel grids were rinsed with PBS 6 x 3 min and then incubated with a 1:100 dilution of gold-conjugated goat anti-rabbit IgG (G7402, Sigma Aldrich) for 2 h at 28°C. The grids were washed and stained with 2% uranyl acetate. Finally, the sections were examined on a transmission electron microscope (H-7650; Hitachi).