Anti mouse cd45 apc mab

Single-cell suspensions of tumor tissue (as described above) and splenic monocyte suspensions (as described above) were blocked with PBS containing 5% BSA for 20 min in room temperature. Subsequently, tissue single-cells were stained with anti-mouse CD45-APC mAb (1:200 dilution; eBioscience™, RRID: AB_469392), anti-mouse CD8-FITC mAb (1:100 dilution; eBioscience™, RRID: AB_464915) and anti-mouse CD31-PE mAb (1:100 dilution; eBioscience™, RRID: AB_465632); Splenic monocytes were stained with anti-mouse CD45-APC mAb (1:200 dilution; eBioscience™, RRID: AB_469392), anti-mouse CD3-FITC mAb (1:100 dilution; eBioscience™, RRID: AB_2572431). After 30 min of staining, the cells were washed twice with PBS and re-suspended in 0.5% BSA (in PBS). Sorting was performed on a Fusion cell sorter (BD FACSAria^TM, USA). In the FSC/SSC-gated monocytes, the CD45⁺CD8⁺ single-cells of tumor tissue were recognized as tumor-infiltrating CD8⁺ T cells, the CD45^-CD31⁺ single-cells of tumor tissue were recognized as vascular endothelial cells, the CD45⁺CD3⁺ splenic monocytes were recognized as splenic T cells.