Coralite 488 labeled goat anti mouse igg

For observation of the localization of Zera-Gn and Zera-Np proteins in Sf9 cells, recombinant baculovirus was first added dropwise to Sf9 cells at an MOI = 0.1; indirect immunofluorescence experiments were conducted as previously reported (18 (link)), and the following antibodies were used in this experiment: mouse anti-His monoclonal antibody (1:500 dilution; Abcam) and the secondary antibodies CoraLite 488-labeled goat anti-mouse IgG (1:500 dilution; Proteintech) and CoraLite 594-labeled goat anti-mouse IgG (1:500 dilution; Proteintech). Laser confocal microscopy (Leica TCS SPE, Wetzlar, Germany) was used to observe the localization of fluorescence. To observe the microstructure of Zera nanoparticles in Sf9 cells, we sent infected Sf9 cells to Xavier for electron microscopy.

Fluorescent tags may affect the immunogenicity of target proteins; thus, we sequenced the recombinant plasmids with the fluorescent tags removed, verified the bacmid sequences by PCR amplification, transfected Sf9 cells and passaged them to P1, P2 and P3 generations with an MOI of 0.5. Viral supernatants were collected, and the baculovirus recombinant proteins rvAc-Gn, rvAc-Np and rvAc- Gn-Np were identified by Western blotting; the primary antibody used was a mouse anti-His monoclonal antibody, and the secondary antibodies were CoraLite 488-labeled goat anti-mouse IgG and CoraLite 594-labeled goat anti-rabbit IgG (1:500 dilution; Proteintech).