Kpl trueblue peroxidase substrate

The SARS-CoV-2 variants, including WHU01, Beta (B.1.351), and Omicron subvariants (BA.1.1, BA.2.3, BA.5, XBB.1, and BQ.1), were preserved in the Guangzhou Customs District Technology Center BSL3 laboratory. The SARS-CoV-2 variants BA.5, XBB.1 and BQ.1 were provided by the Guangdong Provincial Center for Disease Control and Prevention, China. Experiments related to authentic SARS-CoV-2 neutralization were conducted in the BSL3 laboratory of the Guangzhou Customs District Technology Center. Briefly, prediluted antibody or ACE2-Ig proteins were mixed with 100 FFU of SARS-CoV-2 and coincubated at 37°C for 1 h. Then, 50 μL of mixture was transferred to a 96-well plate preseeded with Vero E6 cells. After being maintained at 37°C for 1 h, the cells were covered with MEM containing 1.6% CMC and cultured for 24 h. Cells were then fixed with 4% paraformaldehyde solution, permeabilized with 0.2% Triton X-100, and incubated with anti SARS-CoV/SARS-CoV-2 nucleocapsid rabbit polyclonal antibody (PAb) (Sino Biological; no 40143-T62) and HRP-conjugated goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch; no. 109-035-088). Virus infection foci were stained by KPL TrueBlue peroxidase substrate and read by a CTL ImmunoSpot S6 Ultra reader (Cellular Technology, Ltd.).

The SARS-CoV-2 variants, including WHU01, Beta (B.1.351), and Omicron subvariants (BA.1.1, BA.2.3, BA.5, XBB.1, and BQ.1), were preserved in the Guangzhou Customs District Technology Center BSL3 laboratory. The SARS-CoV-2 variants BA.5, XBB.1 and BQ.1 were provided by the Guangdong Provincial Center for Disease Control and Prevention, China. Experiments related to authentic SARS-CoV-2 neutralization were conducted in the BSL3 laboratory of the Guangzhou Customs District Technology Center. Briefly, prediluted antibody or ACE2-Ig proteins were mixed with 100 FFU of SARS-CoV-2 and coincubated at 37°C for 1 h. Then, 50 μL of mixture was transferred to a 96-well plate preseeded with Vero E6 cells. After being maintained at 37°C for 1 h, the cells were covered with MEM containing 1.6% CMC and cultured for 24 h. Cells were then fixed with 4% paraformaldehyde solution, permeabilized with 0.2% Triton X-100, and incubated with anti SARS-CoV/SARS-CoV-2 nucleocapsid rabbit polyclonal antibody (PAb) (Sino Biological; no 40143-T62) and HRP-conjugated goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch; no. 109-035-088). Virus infection foci were stained by KPL TrueBlue peroxidase substrate and read by a CTL ImmunoSpot S6 Ultra reader (Cellular Technology, Ltd.).