Percp conjugated anti cd3 antibody

The cell surface expression of CD69 and CD25 on the activated human T cells was analyzed by flow cytometry using conjugated corresponding antibodies (anti-human CD69 antibody Cat# 310,902 and anti-human CD25 antibody Cat# 311,702, BioLegend, USA). The VEGFR2-CAR T cells and mock-transduced T cells were co-cultured with HEK-293 and 293-KDR cells at an E:T (effector to target cell) ratio of 1:1. After 24 h, the expression of CD69 and 48 h later, the expression of CD25 were detected by flow cytometry [52 (link), 53 ]. The expression of CD8 on the surface of activated human T cells was evaluated by flow cytometry using anti-CD8 (Cat# 344,702) antibody (BioLegend, USA) followed by staining with conjugated goat anti-mouse secondary antibody (Cat#405,305, BioLegend, USA). VEGFR2-CAR and Mock- transduced T cells (10⁵ cells) were co-cultured with 293-KDR and HEK-293 cells at 1:1 and 3:1 E:T ratios in 96-well plates for 4 h. The cells were then stained using the PE-conjugated anti-CD107a antibody (Cat# 328,607, BioLegend, USA) and PerCP-conjugated anti-CD3 antibody (Cat# 344,813, BioLegend, USA), and after 5 hours were analyzed with flow cytometry.