Apc labeled anti mouse pd l1 antibody

To separate the cells, we utilized the FACSCalibur (BD), followed by the FACSAria (BD). Surface staining assays for flow cytometry and cell sorting assays were conducted. Single cells were suspended in 200 µl of PBS (containing 1% serum) with or without labeled antibodies, following the manufacturer’s protocol. The APC-labeled anti-mouse PD-L1 antibody (Cat#124312; Biolegend) and PE-labeled anti-human PD-L1 antibody (Cat#329706; Biolegend) were employed to stain human and murine PD-L1 in flow cytometry sorting. Isotype PE mouse IgG2b (Cat# 400313, Biolegend) and APC rat IgG2b (Cat# 400612, Biolegend) were used as isotype control antibodies.

Single-cell suspension was obtained as previously described [25 (link)]. The cells were stained with following antibodies; APC-labeled anti-human PD-L1 antibody (#329707, clone 29E.2A3, BioLegend, San Diego, CA, USA), APC-labeled anti-mouse PD-L1 antibody (#124311, clone 10F.9G2, BioLegend), FITC-labeled anti-CD45 (#103107, clone 30-F11, BioLegend), monoclonal PerCP/Cyanine5.5-labeled anti-CD31 (#102419, clone 390, BioLegend), monoclonal PE-labeled anti-CD90.2 (#105307, clone 30-H12, BioLegend), human IgG isotype control antibody (#400322, clone MPC-11, BioLegend), and murine-IgG isotype control antibody (#400612, clone RTK4530, BioLegend). Red blood cell lysis buffer (420302, BioLegend) and Debris Removal Solution (130-109-398, Miltenyi Biotec, Bergisch Gladbach, Germany) were also used. Dead cells (1:1000 dilution) were stained using a Zombie NIR Fixable Viability Kit (423106, BioLegend). Stained cells were analyzed using flow cytometry (FACSLyric; BD Biosciences, Franklin Lakes, NJ, USA), and data were analyzed using the FlowJo software (BD Biosciences).