OriGene overexpression protein microarray chip was immunostained with UltraMAB anti-PDCD1 mouse monoclonal antibody (BD-PE2460). The positive reactive proteins are highlighted with two red arrows in the enlarged subarray. All the positive controls spotted in this subarray are also labeled for clarification (1:100).

Sequential double staining of paraffin human melanoma using b-catenin (red) and PD1 BD-PE2460 (brown). Both abs at 1:800 dilution of 1mg/mL; detection with Polink2 HRP DAB followed by Polink2 Broad AP. Anti-PD1: heat-induced epitope retrieval with Accel; anti-b-catenin: citrate pH6.0. Image shows tumor cells are strongly positve for b-catenin (red) and negative for PD1. The activated T cells (brown) show strong membranous and cytoplasmic staining for PD1 and no staining with b-catenin.

Sequential double staining of paraffin human lung using anti-b-Catenin (red) and anti-PD1 BD-PE2460 (brown). Both antibodies at 1:800 dilution of 1mg/mL. Anti-PD1: heat-induced epitope retrieval with Accel; anti-b-Catenin: citrate pH6.0. Image shows tumor cells are strongly positve for b-catenin (red) and negative for PD1. The arrows point to the activated T cells (brown) showing strong membranous and cytoplasmic staining of PD1 and no staining with b-catenin.

Western blot analysis of extracts (35ug) from PBMCs and PHA-stimulated-PBMCs by using anti-PDCD1 monoclonal antibody (1:250).

Immunohistochemical staining of paraffin-embedded human prostate carcinoma using anti-PD-1 99 mouse monoclonal antibody at 1:800 dilution of 1mg/mL and detection with Polink2 Broad HRP DAB. BD-PE2460 requires heat-induced epitope retrieval with Accel for 3minutes at110C in pressure chamber. The image shows the tumor cells are negative for PD-1 however the activated TCells show strong membranous and cytoplasmic staining.

Immunohistochemical staining of paraffin-embedded human ovarian carcinoma using anti-PD-1 99 mouse monoclonal antibody at 1:800 dilution of 1mg/mL and detection with Polink2 Broad HRP DAB. BD-PE2460 requires heat-induced epitope retrieval with Accel for 3minutes at110C in pressure chamber. The image shows the tumor cells are negative for PD-1 however the activated TCells show strong membranous and cytoplasmic staining.

Immunohistochemical staining of paraffin-embedded human endometrial cancer using anti-PD-1 99 mouse monoclonal antibody at 1:800 dilution of 1mg/mL and detection with Polink2 Broad HRP DAB. BD-PE2460 requires heat-induced epitope retrieval with Accel for 3minutes at110C in pressure chamber. The image shows the tumor cells are negative for PD-1 however the activated TCells show strong membranous and cytoplasmic staining.

Immunohistochemical staining of paraffin-embedded human colon cancer using anti-PD-1 99 mouse monoclonal antibody at 1:800 dilution of 1mg/mL and detection with Polink2 Broad HRP DAB. BD-PE2460 requires heat-induced epitope retrieval with Accel for 3minutes at110C in pressure chamber. The image shows the tumor cells are negative for PD-1 however the activated TCells show strong membranous and cytoplasmic staining.

Western blot of cell lysates (35ug) from 9 different cell lines (1: HepG2, 2: HeLa, 3: SV-T2, 4: A549, 5: COS7, 6: Jurkat, 7: MDCK, 8: PC-12, 9: MCF7). Diluation: 1:500.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PDCD1 (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-PDCD1 antibody (Cat# BD-PE2460)(1:2000).

Western blot of human tissue lysates (15ug) from 10 different tissues (1: Testis, 2: Omentum, 3: Uterus, 4: Breast, 5: Brain, 6: Thyroid, 7: Colon, 8: Spleen 9: Liver, 10: Ovary). Diluation: 1:500.

Immunocytochemistry staining of PDCD1 -stable-expression cells (left) labeling PDCD1 with mouse anti-PDCD1 monoclonal antibody BD-PE2460 (1:900). The rihgt is negative control.

Immunohistochemical staining of paraffin-embedded human melanoma using anti-PD-1 99 mouse monoclonal antibody at 1:800 dilution of 1mg/mL and detection with Polink2 Broad HRP DAB. BD-PE2460 requires heat-induced epitope retrieval with Accel for 3minutes at110C in pressure chamber. The composite image of 3 melanoma shows the tumor cells are negative for PD-1 however the activated TCells show strong membranous and cytoplasmic staining.

Immunohistochemical staining of 3 paraffin-embedded human lung carcinomas using anti-PD1 99 mouse monoclonal antibody at 1:800 requires HIER Accel [OriGene/GBI Labs in Pressure Chamber for 3 minute on high. Detection of primary antibody was achieved with Polink2 Broad HRP DAB [OriGene/GBI Labs D22].

Immunofluorescent staining of PDCD1 -stable-expression cells (left) labeling PDCD1 with mouse monoclonal antibody BD-PE2460 (1:100, green) and nucleus with Hoechst33342 (blue). HEK293T cells serve as negative control (right).

Immunohistochemical staining of paraffin-embedded Human tonsil using anti-PDCD1 mouse monoclonal antibody. (Heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 110°C for 10min, BD-PE2460) (1:1200)