Ina functional flow cytometry test, biotinylated recombinant human B7-H1(10 ng/mL, Catalog # 9049-B7) binds to HEK293 human embryonickidney cell line transfected with human PD-1. Bindingis blocked by 2.5 μg/mL of Mouse Anti-Human PD-1 MonoclonalAntibody (Catalog # MAB10861)
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Please Note: Optimal dilutions should be determined by each laboratory for each application.
are available in the Technical Information section on our website.
Detection of Human PD‑1 by Western Blot.
Western blot shows lysates of HEK293 human embryonic kidney cell line either mock transfected or transfected with human PD-1. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human PD‑1 Monoclonal Antibody (Catalog # MAB10861) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for PD‑1 at approximately 40-80 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) untreated or (B) treated with 5 ng/mL PHA for 2 days were stained with Mouse Anti-Human PD‑1 Monoclonal Antibody (Catalog # MAB10861) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B) and Mouse Anti-Human CD3 epsilon PE‑conjugated Monoclonal Antibody (Catalog # FAB100P). Quadrant markers were set based on control antibody staining (Catalog # MAB002).
PD-1Binding to B7-H1-transfected HEK293 Human Cell Line is Blocked byHuman PD-1 Antibody.
In afunctional flow cytometry test, biotinylated recombinant human B7-H1(10 ng/mL, Catalog # 9049-B7) binds to HEK293 human embryonickidney cell line transfected with human PD-1 (black dotted line). Bindingis completely blocked (orange histogram) by 2.5 μg/mL of Mouse Anti-Human PD-1 MonoclonalAntibody (Catalog # MAB10861). Mouse IgG2B Isotype Control(Catalog # MAB004) at 2.5 μg/mLwas used as a control (blue line). Cells were stained with Streptavidin-APC(Catalog # F0050).
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
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 a type I transmembrane protein belonging to the CD28/CTLA-4 family of immunoreceptors that mediate signals for regulating immune responses (1). Members of the CD28/CTLA-4 family have been shown to either promote T cell activation (CD28 and ICOS) or downregulate T cell activation (CTLA-4 and PD-1) (2). PD-1 is expressed on activated T cells, B cells, myeloid cells, and on a subset of thymocytes. In vitro, ligation of PD-1 inhibits TCR-mediated T cell proliferation and production of IL-1, IL-4, IL-10, and IFN-gamma. In addition, PD-1 ligation also inhibits BCR mediated signaling. PD-1 deficient mice have a defect in peripheral tolerance and spontaneously develop autoimmune diseases (2, 3). Two B7 family proteins, PD-L1 (also called B7-H1) and PD-L2 (also known as B7-DC), have been identified as PD-1 ligands. Unlike other B7 family proteins, both PD‑L1 and PD‑L2 are expressed in a wide variety of normal tissues including heart, placenta, and activated spleens (4). The wide expression of PD-L1 and PD-L2 and the inhibitor effects on PD-1 ligation indicate that PD-1 might be involved in the regulation of peripheral tolerance and may help prevent autoimmune diseases (2). The human PD-1 gene encodes a 288 amino acid (aa) protein with a putative 20 aa signal peptide, a 148 aa extracellular region with one immunoglobulin-like V‑type domain, a 24 aa transmembrane domain, and a 95 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 60% aa sequence identity (4).
Ishida, Y. et al. (1992) EMBO J. 11:3887.
Nishimura, H. and T. Honjo (2001) Trends in Immunol. 22:265.
Latchman, Y. et al. (2001) Nature Immun. 2:261.
Carreno, B.M. and M. Collins (2002) Annu. Rev. Immunol. 20:29.
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