Fitc goat anti mouse igg antibody

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).

Primary antibodies for Flow Cytometry: mouse anti-latrophilin 2/AGRL2/LPHN2 (1 µg/10⁶ cells; R&D Systems, #MAB105881-SP); mouse anti-glypican 4 (1 µg/10⁶ cells; R&D Systems, MAB9195-SP); mouse anti-JAM3 (1 µg/10⁶ cells; R&D SYSTEMS, MAB11891-SP); PE-conjugated anti-L1CAM, (1:20; Miltenyi Biotec, Bergisch Gladbach, Germany; #130-100-691); PE-conjugated anti-FGFR4 (1:100; Biolegend, London, UK; #324306). Secondary antibodies: FITC Goat anti-mouse IgG antibody (1:500; BioLegend, #405305). Isotype controls: mouse FITC-conjugated IgG1, κ Isotype Ctrl Antibody (1:100; BioLegend, #400108); rabbit PE-conjugated (1:100; R&D Systems, #AB-105-C).