Human PD-1 Antibody

Recombinant Monoclonal Antibody
    
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  • Species Reactivity
    Human
  • Specificity
    Detects human PD-1 in direct ELISAs.
  • Source
    Recombinant Monoclonal Mouse IgG1 Clone # 913429
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human PD-1
    Met1-Gln167
    Accession # Q15116
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Blockade of Receptor-ligand Interaction
    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)
  • CyTOF-ready
    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. General Protocols are available in the Technical Information section on our website.
Data Examples
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).
Blockade of Receptor-ligand Interaction
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
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
    Reconstitution Buffer Available
  • Shipping
    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.
Background: PD-1

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).

  • References:
    1. Ishida, Y. et al. (1992) EMBO J. 11:3887.
    2. Nishimura, H. and T. Honjo (2001) Trends in Immunol. 22:265.
    3. Latchman, Y. et al. (2001) Nature Immun. 2:261.
    4. Carreno, B.M. and M. Collins (2002) Annu. Rev. Immunol. 20:29.
  • Long Name:
    Programmed Death-1
  • Entrez Gene IDs:
    5133 (Human); 18566 (Mouse); 301626 (Rat); 486213 (Canine); 102123659 (Cynomolgus Monkey)
  • Alternate Names:
    CD279 antigen; CD279; hPD-1; PD-1; PD1hPD-l; PDCD1; programmed cell death 1; programmed cell death protein 1; Protein PD-1; SLEB2
Related Research Areas

FAQs

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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 149  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Reconstitution Buffers
Description Application Cat# Citations Images  

Reconstitution Buffer 1 (PBS)

RB01
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 33  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 12  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow , IHC , ICC NL007 10
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow , IHC , ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 4  
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Mouse IgG Antibody

WB AF007 4
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow , IHC , ICC NL009 5
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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