Anti PD-1  

Figure 1 KO Validation in HeLa Cells
Loading: 10 μg of HeLa WT cell lysates or PD-1 KO cell lysates. Antibodies: PD-1, 4065 (4 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 2 KD Validation in HeLa Cells
Loading: 10 μg of HeLa WT cell lysates or PD-1 KD cell lysates. Antibodies: PD-1, 4065 (4 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 3 Western Blot Validation in Human and Mouse Cell Lines
Loading: 15 μg of lysates per lane.Antibodies: PD-1 4065 (4 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 4 Western Blot Validation in THP-1 Cell Lysate in the (A) absence and (B) presence of blocking peptide
Loading: 15 μg of lysates per lane.Antibodies: PD-1, 4065 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 5 Western Blot Validation in Rat Thymus Cell Lysate
Loading: 15 μg of lysates per lane.Antibodies: PD-1 4065 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 6 Immunohistochemistry Validation of PD-1 in Human Tonsil Tissue
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PD-1 antibody (4065) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4℃. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Figure 6 Immunohistochemistry Validation of PD-1 in Human Tonsil Tissue
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PD-1 antibody (4065) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4℃. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Figure 8 Immunohistochemistry Validation of PD-1 in Human Brain Tissue
Immunohistochemical analysis of paraffin-embedded human brain tissue using anti-PD-1 antibody (4065) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4℃. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.