Anti sars cov 2 s protein

Protein concentrations of the extracts were assayed with Bradford reagent (Bio-Rad), and equal amounts of cell extracts were separated by SDS–12% PAGE and transferred onto a nitrocellulose membrane (Sigma). Samples were then analyzed by Western blotting using the desired antibodies anti-SARS-CoV-2 S protein (catalog number 40591-T62, Sino Biologicals), anti-SARS-CoV-2 N protein (catalog number 40143-MM05, Sino-Biologicals), and immunized mouse sera followed by the respective secondary antibodies (horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG; Sigma). Antibody complexes were detected using Immobilon Western systems (Millipore).

Protein concentrations of the extracts were assayed with Bradford reagent (Bio-Rad), and equal amounts of cell extracts were separated by SDS–12% PAGE and transferred onto a nitrocellulose membrane (Sigma). Samples were then analyzed by Western blotting using the desired antibodies anti-SARS-CoV-2 S protein (catalog number 40591-T62, Sino Biologicals), anti-SARS-CoV-2 N protein (catalog number 40143-MM05, Sino-Biologicals), and immunized mouse sera followed by the respective secondary antibodies (horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG; Sigma). Antibody complexes were detected using Immobilon Western systems (Millipore).

Cells were fixed with 4% (w/v) paraformaldehyde in PBS at RT for 15 min and blocked in PBS buffer containing 10% donkey serum and 0.3% Triton X-100 (Sigma, T8787) for 1 h at RT, followed by incubation with the primary antibodies at 4 °C overnight with 5% donkey serum and 0.15% Triton X-100. Nuclei were counterstained with DAPI DNA dye (CST, 4083, 1:1000) at RT for 10 min and mounted on glass slides. Images were taken using a PerkinElmer High Content Analysis System Operetta CLS and processed using Harmony 4.9 software. The following primary antibodies were used for immunofluorescence: anti-SARS-CoV-2 S protein (Sino Biological, Rabbit, T62, 1:500), anti-METTL14 and anti-ALKBH5 (Proteintech, 26158-1-AP and 16837-1-AP, Rabbit, 1:500).