Fitc anti rabbit igg h l

For FCGRs detection, primary B cells, monocytes, and macrophages were incubated with fluorochrome-labeled antibodies specific for humans before fixation: FITC mouse anti-human CD32 (552883; BD Pharmingen, San Diego, CA, USA), APC mouse anti-human CD64 (561189; BD Pharmingen, San Diego, CA, USA ), CD16 Rabbit PAb (16559-1-AP; Proteintech,--). FITC-anti-Rabbit IgG (H+L) (SA00003-2; Proteintech, Chicago, IL, USA) was used as the secondary antibody for CD16.
For SARS-CoV-2 infected primary immune cells, surface staining was conducted before fixation with AF700-anti-CD45 (368514; BioLegend, San Diege, CA, USA), BV650-anti-CD11b (101239; BioLegend, San Diege, CA, USA), PE-anti-CD68 (333808; Biolegend, San Diege, CA, USA), and Percp Cy5.5-anti-CD14 (367110; Biolegend, San Diege, CA, USA). Antibody stained cells were fixed overnight with 4% PFA at 4 °C and taken out of the BSL3 lab for downstream analysis. Cells were stained further with in house made SARS-CoV NP pAb (1:500) at 4 °C for 30 min after permeabilization. Then, cells were stained with FITC-anti-Rabbit IgG (H + L) (SA00003-2; Proteintech, Chicago, IL, USA) at room temperature for 30 min.

For surface staining, PBCs were incubated with fluorochrome-labeled antibodies specific for humans before fixation: AF-700-anti-CD45 (2D1), percp-anti-CD19 (HIB19), APC/CY7-anti-CD3 (UCHT1), BV510-anti-CD4 (OKT4) and percp/Cy5.5-anti-CD8a (HTT8a). Antibody-stained PBCs were fixed overnight with 4% PFA at 4 °C and taken out of BSL3 lab for downstream analysis. Cells were stained further with in-house-made SARS-CoV-1 NP pAb (1:500) at 4 °C for 30 min after permeabilization. Then cells were stained with FITC-anti-Rabbit IgG (H + L) at room temperature for 30 min. AF-700-anti-CD45 (2D1), APC/CY7-anti-CD3 (UCHT1), BV510-anti-CD4, and percp/Cy5.5-anti-CD8a antibodies were purchased from Biolegend and all were used at 1:100. FITC-anti-Rabbit IgG (H + L) was from Proteintech (SA00003-2).