Human PD-1 Antibody

(12 citations)
(3 Reviews)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human PD-1 in ELISAs and Western blots. In sandwich ELISAs, less than 2% cross-reactivity with recombinant mouse PD‑1 and less than 0.2% cross-reactivity with recombinant human (rh) CD28, rhICOS, and rhCTLA‑4 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human PD-1
    Leu25-Gln167
    Accession # Q8IX89
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5-2 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Blockade of Receptor-ligand Interaction
    In a functional ELISA, 3-12 µg/mL of this antibody will block 50% of the binding of 500 ng/mL of Recombinant Human B7-H1 Fc Chimera (Catalog # 156-B7) to immobilized Recombinant Human PD-1 Fc Chimera (Catalog # 1086-PD) coated at 1 µg/mL (100 µL/well). At 30 μg/mL, this antibody will block >90% of the binding.
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
    • Human PD-1 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Human PD‑1 Antibody (Catalog # AF1086)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human PD‑1 Biotinylated Antibody (Catalog # BAF1086)
  • ELISA Standard
     
    Recombinant Human PD-1 Fc Chimera Protein, CF (Catalog # 1086-PD)
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 lysate of human thymus tissue. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for PD‑1 at approximately 40-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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 0.5 µg/mL of Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). 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 treated with PHA by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either (A) untreated or (B) treated with 5 μg/mL PHA overnight were stained with Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.
Immunohistochemistry
PD‑1 in Human Lymph Node. PD‑1 was detected in immersion fixed paraffin-embedded sections of human lymph node using Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) at 3 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 lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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 down-regulate 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 immuno-receptor
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
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Immune Cell Dynamics in Rhesus Macaques Infected with a Brazilian Strain of Zika Virus
    Authors: ELV Silveira, KA Rogers, S Gumber, P Amancha, P Xiao, SM Woollard, SN Byrareddy, MM Teixeira, F Villinger
    J. Immunol., 2017;0(0):.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Tissue
    Application: IHC
  2. Increased expression of programmed cell death protein 1 on NK cells inhibits NK-cell-mediated anti-tumor function and indicates poor prognosis in digestive cancers
    Authors: Y Liu, Y Cheng, Y Xu, Z Wang, X Du, C Li, J Peng, L Gao, X Liang, C Ma
    Oncogene, 2017;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  3. Vaccine Induction of Lymph Node-Resident Simian Immunodeficiency Virus Env-Specific T Follicular Helper Cells in Rhesus Macaques.
    Authors: Vargas-Inchaustegui D, Demers A, Shaw J, Kang G, Ball D, Tuero I, Musich T, Mohanram V, Demberg T, Karpova T, Li Q, Robert-Guroff M
    J Immunol, 2016;196(4):1700-10.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Tissue
    Application: IHC Fluorescence
  4. PD-1 Blockade with Pembrolizumab in Advanced Merkel-Cell Carcinoma
    Authors: PT Nghiem, S Bhatia, EJ Lipson, RR Kudchadkar, NJ Miller, L Annamalai, S Berry, EK Chartash, A Daud, SP Fling, PA Friedlande, HM Kluger, HE Kohrt, L Lundgren, K Margolin, A Mitchell, T Olencki, DM Pardoll, SA Reddy, EM Shantha, WH Sharfman, E Sharon, She
    N Engl J Med, 2016;0(0):.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
  5. Plasmacytoid dendritic cells promote HIV-1-induced group 3 innate lymphoid cell depletion.
    Authors: Zhang Z, Cheng L, Zhao J, Li G, Zhang L, Chen W, Nie W, Reszka-Blanco N, Wang F, Su L
    J Clin Invest, 2015;125(9):3692-703.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  6. Soluble co-signaling molecules predict long-term graft outcome in kidney-transplanted patients.
    Authors: Melendreras S, Martinez-Camblor P, Menendez A, Bravo-Mendoza C, Gonzalez-Vidal A, Coto E, Diaz-Corte C, Ruiz-Ortega M, Lopez-Larrea C, Suarez-Alvarez B
    PLoS ONE, 2014;9(12):e113396.
    Species: Human
    Sample Type: Serum
    Application: ELISA Development
  7. Early lymphoid responses and germinal center formation correlate with lower viral load set points and better prognosis of simian immunodeficiency virus infection.
    Authors: Hong J, Amancha P, Rogers K, Courtney C, Havenar-Daughton C, Crotty S, Ansari A, Villinger F
    J Immunol, 2014;193(2):797-806.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Tissue
    Application: IF
  8. CD4 T follicular helper cell dynamics during SIV infection.
    J. Clin. Invest., 2012;122(9):3281-94.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  9. Modulation of T-cell activation by malignant melanoma initiating cells.
    Authors: Schatton T, Schutte U, Frank NY, Zhan Q, Hoerning A, Robles SC, Zhou J, Hodi FS, Spagnoli GC, Murphy GF, Frank MH
    Cancer Res., 2010;70(2):697-708.
    Species: Human
    Sample Type: Whole Tissue
    Application: IF
  10. Early resolution of acute immune activation and induction of PD-1 in SIV-infected sooty mangabeys distinguishes nonpathogenic from pathogenic infection in rhesus macaques.
    Authors: Estes JD, Gordon SN, Zeng M, Chahroudi AM, Dunham RM, Staprans SI, Reilly CS, Silvestri G, Haase AT
    J. Immunol., 2008;180(10):6798-807.
    Species: Primate - Cercocebus torquatus (Sooty Mangabey)
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  11. Soluble PD-1 rescues the proliferative response of simian immunodeficiency virus-specific CD4 and CD8 T cells during chronic infection.
    Authors: Onlamoon N, Rogers K, Mayne AE, Pattanapanyasat K, Mori K, Villinger F, Ansari AA
    Immunology, 2008;124(2):277-93.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Cells
    Application: Neut
  12. Aberrant regulation of synovial T cell activation by soluble costimulatory molecules in rheumatoid arthritis.
    Authors: Wan B, Nie H, Liu A, Feng G, He D, Xu R, Zhang Q, Dong C, Zhang JZ
    J. Immunol., 2006;177(12):8844-50.
    Species: Human
    Sample Type: Serum
    Application: ELISA Development
Expand to show all 12 Citations
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Average Rating: 5 (Based on 3 reviews)

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We have 3 reviews tested in 3 applications: Simple Western, ELISA, Flow Cytometry.

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Images Ratings Applications Species Reviewed By Date Details
  Excellent
 Simple Western Human Anonymous 11/24/2017
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Summary

ApplicationSimple Western
Sample Tested2120Ep human embryonal carcinoma cell line
SpeciesHuman
  Excellent
 ELISA Human Anonymous 09/29/2017
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Summary

ApplicationELISA
Sample TestedHuman recombinant test antibody
SpeciesHuman
Flow Cytometry Human PD-1 Antibody AF1086
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Excellent
 Flow Human Anonymous 03/14/2017
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Flow Cytometry Human PD-1 Antibody AF1086
Flow Cytometry: Human PD-1 Antibody [AF1086]

Summary

ApplicationFlow Cytometry
Sample TestedPeripheral blood mononuclear cells (PBMCs)
SpeciesHuman

Other Experimental Details

Other Experimental Details10^6 Human PBMCs were probed with 0.5 ug of Goat anti-Human PD-1 antibody (red), or Goat Isotype Control IgG (black), followed by PE-conjugated anti-Goat secondary antibody.

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