Dylight 488 conjugated goat anti rabbit igg secondary antibody

IBRS-2 cells were infected with SVA strain A/ZJ/2015 (MOI = 0.5) when they reached 70% confluence. Cells were inoculated at 37°C with 5% CO₂ over a 1-h period, the medium was replaced with fresh DMEM, and continued incubation. DMEM was removed at 0, 6, 9, and 12 hpi, and cells were fixed with cold methanol for 10 min. Samples were washed with PBS, blocked with 1% bovine serum albumin (Solarbio) for 30 min, and SVA proteins were detected using rabbit polyclonal anti-SVA-VP1 primary antibody (prepared and stored in our laboratory) and Dylight 488-conjugated goat anti-rabbit IgG secondary antibody (Abbkine, Wuhan, China). Cell nuclei were stained for 20 min with 10 μg/ml Hoechst 33342 (Solarbio), and immunofluorescence microscopy images were captured using a confocal laser scanning microscope (Leica, Wetzlar, Germany).

Lipopolysaccharide (LPS), 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2Htetrazolium bromide (MTT), and dimethylsulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Griess Reagent System was obtained from Promega Corporation (Madison, WI, USA). The commercial-specific complete Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin, and streptomycin were purchased from GIBCO (Carlsbad, CA, USA). The ECL kit was provided from GE Healthcare. 4′,6-diamidino-2-phenylindole (DAPI) was obtained from Solarbio (Beijing, China); primary and secondary antibodies used for western blot were purchased from Santa Cruz Biotechnology Inc (Santa Cruz, CA, USA). The anti-NF-κB p65 antibody was obtain from Proteintech Group (Rosemont, IL, USA); Dylight 488 conjugated Goat Anti-Rabbit IgG secondary antibody was purchased from Abbkine (Redlands, CA, USA). The commercial ELISA kit for detecting IL-1β was obtained from Beyotime Biotechnology (Nanjing, China). Those chemicals not mentioned in present research were of analytic grade.