PD-L1 Luminex ELISA with 2C7 Capture (BD-PE3486) and 9E12 Detection Antibodies. Substrate used: Recombinant Human PD-L1

Detection of PDL1 neutralizing antibody using MACS column. GFP+/PDL1+ 293T cells (co-transfected with PDL1 and GFP plasmid (PS10010) were incubated with either PDL1 antibody BD-PE3486 (red), non-specific antibody (green), isotype control (blue) or PBS (black) and then mixed with PD1+ 293T cells linked with magnetic-beads. The mixed cells were pulled down using MACS column (Miltenyi Biotec) and analysed by Flow Cytometry. GFP+/PDL1+ cells would not be collected if PD1/PDL1 interaction is neutralized by the tested antibody (1:50).

Detection of PDL1 neutralizing antibody using MACS column. PD1+ 293T cells linked with magnetic-beads and GFP+/PDL1+ 293T cells co-transfected with PDL1 and GFP plasmids were mixed together and incubated with PDL1 antibody BD-PE3486 (red), mouse IgG isotype control (blue) or PBS (black). The mixed cells were pulled down using MACS column (Miltenyi Biotec) and analysed by Flow Cytometry. GFP+/PDL1+ cells would be collected if PD1/PDL1 interaction is not neutralized by the tested antibody.

Immunocytochemistry staining of CD274 stable expression cells using anti-CD274 mouse monoclonal antibody (BD-PE3486) (Left). The right is negative control (1:5000).

PD-L1 ELISA with 2C7 Capture (BD-PE3486) and 9E12 Detection Antibodies. Substrate used: Recombinant Human PD-L1

Anti-CD274 mouse monoclonal antibody (BD-PE3486) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY CD274 .

Flow cytometric analysis of living PBMCs treated with 10ug/ml PHA for 72h (Right)/untreated (Left) using anti-PDL1 antibody (BD-PE3486) (1:100).

Flow cytometric analysis of living PBMCs treated with 10ug/ml PHA for 72h (Right) using anti-PDL1 antibody (BD-PE3486). Cells incubated with a non-specific antibody (Left) were used as isotype control (1:100).

Flow cytometric Analysis of HCC78 cells, using anti-PDL1 antibody (BD-PE3486), (Red), compared to isotype control, (green), and negative control (PBS), (Blue) (1:100).

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CD274 (Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CD274(Cat# BD-PE3486). Positive lysates (100ug) and (20ug) can be purchased separately from OriGene.