Fitc anti mouse cd19 antibody

Mouse spleen cells were harvested and divided into three parts. The first portion of the spleen cells was treated with 100 μL of 20 μg/mL FITC anti-mouse CD3 antibody (BioLegend, San Diego, CA, USA), 100 μL of 12.5 μg/mL PerCP anti-mouse CD4 antibody (BioLegend), 100 μL of 25 μg/mL APC anti-mouse CD25 antibody (BioLegend), and 100 μL of 20 μg/mL PE anti-mouse Foxp3 antibody (BioLegend) at 25 °C for 30 min, washed, and then incubated with 100 μL of 10 μg/mL FITC-conjugated goat anti-rat IgG (BioLegend) at 25 °C for 30 min in the dark. The second portion of the spleen cells was treated with 100 μL of 20 μg/mL FITC anti-mouse CD3 antibody (BioLegend), 100 μL of 12.5 μg/mL APC anti-mouse CD8 antibody (BioLegend), and 100 μL of 12.5 μg/mL PE anti-mouse CD28 antibody (BioLegend) using the same protocol as above. The third portion of spleen cells was treated with 100 μL of 20 μg/mL FITC anti-mouse CD19 antibody (BioLegend) and 100 μL of 50 μg/mL PE anti-mouse CD80 antibody (BioLegend) using the same protocol as above. After incubation, the cells were washed and resuspended in 0.5 mL of PBS/2% paraformaldehyde, and then quantified using flow cytometry (BD Calibur™, San Jose, CA, USA).

Mouse spleen cells were harvested and divided into three parts. The first part of the spleen cells were treated with 100μL of 20μg/mL FITC anti-mouse CD3 antibody (BioLegend, San Diego, USA), 100μL of12.5μg/mL Percp anti-mouse CD4 antibody (BioLegend) and 100μL of 12.5μg/mL APC anti-mouse CD8 antibody (BioLegend) at 25°C for 30 min, washed, and followed by incubation with 100μL of 10μg/mL FITC-conjugated goat anti-rat IgG (BioLegend) and incubated at 25°C for 30 min in the dark. The second part of the spleen cells were treated with 100μL of 20μg/mL FITC anti-mouse CD3 antibody (BioLegend), 100μL of 12.5μg/mL Percp anti-mouse CD4 antibody (BioLegend),100μL of 25μg/mL APC anti-mouse CD25 antibody (BioLegend) and 100μL of 20 μg/mL PE anti- mouse FOXP3 (BioLegend) using the same protocol as above. The third part of spleen cells were treated with 100μL of 20μg/mL FITC anti-mouse CD19 antibody (BioLegend) and 100μL of 50μg/mL PE anti-mouse CD80 antibody (BioLegend) using the same protocol as above. After incubation, the cells were washed and resuspended in 0.5 mL of PBS/2% paraformaldehyde, then quantified by flow cytometry (BD Calibur™).

Mouse blood cells were harvested and divided into three parts. First, spleen cells were treated with 100 μL of 20 μg/mL FITC anti-mouse CD3 antibody (100306, BioLegend, San Diego, USA), 100 μL of 12.5 μg/mL Percp anti-mouse CD4 antibody (100407, BioLegend), and 100 μL of 12.5 μg/mL APC anti-mouse CD8 antibody (100732, BioLegend) at 25°C for 30 min, washed, then incubated with 100 μL of 10 μg/mL FITC goat anti-mouse IgG antibody (405305, BioLegend) at 25°C for 30 min in the dark. Second, blood cells were treated with 100 μL of 12.5 μg/mL Percp anti-mouse CD4 antibody (BioLegend), 100 μL of 25 μg/mL APC anti-mouse CD25 antibody (101910, BioLegend), and 100 μL of 12.5 μg/mL PE anti-mouse Foxp3 antibody (320008, BioLegend) via the same protocol described above. Third, blood cells were treated with 100 μL of 20 μg/mL FITC anti-mouse CD19 antibody (115506, BioLegend) using the same protocol described above. After incubation, cells were washed and resuspended in 0.5 mL of PBS/2% paraformaldehyde and quantified by flow cytometry (BD Calibur^TM).