Detection of Mouse PD‑1 by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line mock transfected or transfected with mouse PD-1. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PD‑1 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
PD‑1 in Mouse Thymus. PD‑1 was detected in perfusion fixed frozen sections of mouse thymus using 15 µg/mL Mouse PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1021) overnight at 4 °C. Tissue was stained (red) and counterstained (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Programmed Death-1 (PD-1) is type I transmembrane protein belonging to the CD28/CTLA-4 family of immunoreceptors that mediate signals for regulating immune responses (1). Other members of this family include CD28, CTLA-4, and ICOS (2-4). PD-1 is most closely related to CTLA-4 and shares approximately 24% amino acid (aa) sequence identity. The mouse PD-1 gene encodes a 288 aa protein with a putative 20 aa signal peptide, a 149 aa extracellular region with one immunoglobulin-like V-type domain, a 21 aa transmembrane domain, and a 98 aa cytoplasmic region. The cytoplasmic tail contains two tyrosine residues that form the immunoreceptor tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM) that are important in mediating PD-1 signaling. Mouse and human PD-1 share approximately 69% aa sequence identity. Two B7 family proteins, PD-L1 (also called B7-H1) and PD-L2, have been identified as PD-1 ligands (5, 6). PD-1 is expressed on activated T cells, B cells, myeloid cells, and on a subset of thymocytes. PD-1 deficient mice have a defect in peripheral tolerance and spontaneously develop autoimmune diseases. Binding of PD-1 to PD-L1 or PD-L2 results in the inhibition of TCR-mediated proliferation and cytokine production as well as BCR-mediated signaling. PD-1 likely has an inhibitory role in regulating immune responses (1-4).
Ishida, Y. et al. (1992) EMBO J. 11:3887.
Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116.
Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
Nishimura, H. and T. Honjo (2001) Trends in Immunol. 22:265.
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