PD-1 Rabbit Monoclonal Antibody(ARA754)

All lanes: Anti-PD-1 antibody at 1:5,000 dilution
Predicted MW: 31 kDa
Observed MW: 40-50 kDa

Lane 1: 293 Overexpression of HuPD-1
Lane 2: 293 w/o HuPD-1

Lysates at 5 µg per lane
2nd Ab:
GAR HRP(H+L) 1:1,000
Exposure: 120s

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung tonsil tissue labelling PD-1 with ARA754 at 1:200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0.

ARA754 staining PD-1 in 293 cells transfected with PD-1 gene by IF/ICC (immunofluorescence/immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% goat serum for half an hour at room temperature. Samples were incubated with primary antibody (1:10,000) at 4°C. An Alexa Fluor^® 488-conjugated Goat Anti-Rabbit IgG polyclonal was used as the secondary antibody (1:500). DAPI (blue) was used as the nuclear counter stain.
Control: PBS and secondary antibody, An Alexa Fluor^® 488-conjugated Goat Anti-Rabbit IgG (1:500).

Overlay histogram showing 293 cells transfected with PD-1 gene stained with ARA754 (Blue). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% TritonX-100 for 15 min. The cells were then incubated in the antibody (ARA754, 1:200 dilution) in 1x PBS/1% BSA for 30 min at room temperature. The secondary antibody used was a Goat Anti-Rabbit Alexa Fluor^® 488 (IgG H+L) at 1:2,000 dilution for 20 min at room temperature. Unlabelled sample (Red) was used as a control.

PD-1 was immunoprecipitated from 0.2mg of 293 whole cell lysate transfected with PD-1 gene with ARA754 at 1:50 dilution.
2nd Ab:
GAR HRP for IP 1:500
Lane 1: ARA754 IP in 293 whole cell lysate transfected with PD-1 gene
Lane 2: PBS instead of ARA754 in 293 whole cell lysate transfected with PD-1 gene
Lane 3: 293 whole cell lysate transfected with PD-1 gene, 2 µg (input)
Exposure: 10s