Mouse PD-1 Antibody

PD‑1 in Mouse Spleen.

PD-1 was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse PD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in splenocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

PD‑1 in Mouse Thymus.

PD-1 was detected in perfusion fixed paraffin-embedded sections of mouse thymus using Goat Anti-Mouse PD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Mouse PD-1 by Western Blot

Expression and glycosylation of Siglec-1 counter-receptors on resting and activated Tregs.(A) Eight Siglec-1 counter receptors were randomly selected for cytometry analysis. All experiments were performed three times and similar results were observed. (B) Normalized mRNA counts of Siglec-1 counter receptors were obtained from Treg RNA-Seq data. The result is visualized in a volcano plot. Only a small subset of counter-receptors showed strongly increased gene expression following Treg activation. The counter-receptors selected for flow cytometry analysis are highlighted in red. (C) Western blot showing that CD48 had a higher molecular weight in activated Tregs. (D) Western blot of PD-1 affinity-purified from resting and activated Tregs. PD 1 showed a dramatic decrease of molecular weight after PNGase F digestion (D), PD-1 from activated Tregs had higher molecular weight (E) and following sialidase treatment, PD-1 from resting and activated Tregs migrated at a slightly reduced molecular weight indicating the presence of sialic acids (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35224210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse PD-1 by Western Blot

Expression and glycosylation of Siglec-1 counter-receptors on resting and activated Tregs.(A) Eight Siglec-1 counter receptors were randomly selected for cytometry analysis. All experiments were performed three times and similar results were observed. (B) Normalized mRNA counts of Siglec-1 counter receptors were obtained from Treg RNA-Seq data. The result is visualized in a volcano plot. Only a small subset of counter-receptors showed strongly increased gene expression following Treg activation. The counter-receptors selected for flow cytometry analysis are highlighted in red. (C) Western blot showing that CD48 had a higher molecular weight in activated Tregs. (D) Western blot of PD-1 affinity-purified from resting and activated Tregs. PD 1 showed a dramatic decrease of molecular weight after PNGase F digestion (D), PD-1 from activated Tregs had higher molecular weight (E) and following sialidase treatment, PD-1 from resting and activated Tregs migrated at a slightly reduced molecular weight indicating the presence of sialic acids (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35224210), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse PD-1 by Western Blot

Expression and glycosylation of Siglec-1 counter-receptors on resting and activated Tregs.(A) Eight Siglec-1 counter receptors were randomly selected for cytometry analysis. All experiments were performed three times and similar results were observed. (B) Normalized mRNA counts of Siglec-1 counter receptors were obtained from Treg RNA-Seq data. The result is visualized in a volcano plot. Only a small subset of counter-receptors showed strongly increased gene expression following Treg activation. The counter-receptors selected for flow cytometry analysis are highlighted in red. (C) Western blot showing that CD48 had a higher molecular weight in activated Tregs. (D) Western blot of PD-1 affinity-purified from resting and activated Tregs. PD 1 showed a dramatic decrease of molecular weight after PNGase F digestion (D), PD-1 from activated Tregs had higher molecular weight (E) and following sialidase treatment, PD-1 from resting and activated Tregs migrated at a slightly reduced molecular weight indicating the presence of sialic acids (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35224210), licensed under a CC-BY license. Not internally tested by R&D Systems.