Human PD-1 Antibody Summary
Accession # Q15116
This antibody functions as an ELISA capture antibody when paired with Mouse Anti-Human PD‑1 Monoclonal Antibody(Catalog # MAB10864).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human PD-1 DuoSet ELISA Kit (Catalog # DY1086) for convenient development of a sandwich ELISA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human PD‑1 by Western Blot. Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with human PD-1. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human PD-1 Monoclonal Antibody (Catalog # MAB10863) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for PD-1 at approximately 40 kDa and 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of PD‑1 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either (A) treated with 5 ng/mL PHA for 2 days or (B) untreated, were stained with Rabbit Anti-Human PD-1 Monoclonal Antibody (Catalog # MAB10863) followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111) and Mouse Anti-Human CD3 epsilon PE-conjugated Monoclonal Antibody (Catalog # FAB100P). Quadrant markers were set based on control antibody staining (Catalog # MAB1050). View our protocol for Staining Membrane-associated Proteins.
Human PD‑1 ELISA Standard Curve. Recombinant Human PD-1 protein was serially diluted 2-fold and captured by Rabbit Anti-Human PD-1 Monoclonal Antibody (Catalog # MAB10863) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human PD-1 Monoclonal Antibody(Catalog # MAB10864) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Preparation and Storage
- 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 receptor (PD-1), also known as CD279, is type I transmembrane protein belonging to the CD28 family of immune regulatory receptors (1). Other members of this family include CD28, CTLA-4, ICOS, and BTLA (2-5). Mature human PD-1 consists of a 148 amino acid (aa) extracellular region (ECD) with one immunoglobulin-like V-type domain, a 24 aa transmembrane domain, and a 95 aa cytoplasmic region. The human PD-1 ECD shares 65% aa sequence identity with the mouse PD-1 ECD. 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 for mediating PD-1 signaling. PD-1 acts as a monomeric receptor and interacts in a 1:1 stoichiometric ratio with its ligands PD-L1 (B7-H1) and PD-L2 (B7-DC) (6, 7). PD‑1 is expressed on activated T cells, B cells, monocytes, and dendritic cells while PD-L1 expression is constitutive on the same cells and also on nonhematopoietic cells such as lung endothelial cells and hepatocytes (8, 9). Ligation of PD-L1 with PD-1 induces co-inhibitory signals on T cells promoting their apoptosis, anergy, and functional exhaustion (10). Thus, the PD-1: PD-L1 interaction is a key regulator of the threshold of immune response and peripheral immune tolerance (11). Finally, blockade of the PD-1: PD-L1 interaction by either antibodies or genetic manipulation accelerates tumor eradication and shows potential for improving cancer immunotherapy (12, 13, 14).
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