Mouse anti his monoclonal antibody

Attenuated live IBDV vaccine was from Ringpu (Baoding, China). Mouse anti-His monoclonal antibody and goat anti-mouse IgG-AP antibody were from Abcam (Cambridge, MA, USA). Recombinant IBDV VP2 protein was prepared in our laboratory as previously described [17 (link)].

The larger molecular weight of the fluorescent tags may affect the spatial structure and function of proteins, which further affects the immune effect. Therefore, we modified the two vectors to produce proteins without fluorescent tags, named dual-Zera-Gn and dual-Zera-Np, and we prepared these nonfluorescent recombinant baculoviruses using the abovementioned method. Zera-Gn and Zera-Np nanoparticles were purified by sucrose density gradient centrifugation and identified by western blotting. The primary antibody was a mouse anti-His monoclonal antibody (1:5000 dilution; Abcam); the secondary antibody was a rabbit anti-mouse monoclonal antibody (1:5000 dilution; Proteintech).

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.