Human PD‑1 Antibody

Catalog # AF1086 | R&D Systems, Inc. a Bio-Techne Brand
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AF1086
AF1086-SP
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Key Product Details

Species Reactivity

Human

Applications

Blockade of Receptor-ligand Interaction, CyTOF-ready, ELISA Capture (Matched Antibody Pair), Flow Cytometry, Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Summary for Human PD‑1 Antibody

Immunogen

Mouse myeloma cell line NS0-derived recombinant human PD-1
Leu25-Gln167
Accession # Q8IX89

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human PD‑1 Antibody

Detection of Human PD‑1 antibody by Western Blot.

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 antibody by Western Blot.

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 antibody in Human PBMCs treated with PHA antibody by Flow Cytometry.

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 CD3e 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.
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Applications for Human PD‑1 Antibody

Application
Recommended Usage

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.

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells (PBMCs) treated with PHA

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymph node

Western Blot

0.5-2 µg/mL
Sample: Human thymus tissue and HEK293 human embryonic kidney cell line transfected with human PD-1

Human PD-1 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human PD‑1 Biotinylated Antibody (Catalog # BAF1086)
  • Standard: Recombinant Human PD-1 Fc Chimera Protein, CF (Catalog # 1086-PD)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 6 reviews rated 4.5 using AF1086 in the following applications:

Formulation, Preparation and Storage

Purification

Antigen Affinity-purified

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in 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

Alternate Names

CD279, PD1, PDCD1, SLEB2

Entrez Gene IDs

486213 (Canine); 102123659 (Cynomolgus Monkey); 5133 (Human); 18566 (Mouse); 100533201 (Porcine); 301626 (Rat)

Gene Symbol

PDCD1

UniProt

Q8IX89

Product Documents for Human PD‑1 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human PD‑1 Antibody

For research use only

Citations for Human PD‑1 Antibody (34)

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Customer Reviews for Human PD‑1 Antibody (6)

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  • Human PD-1 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Melanoma tissue
    Species: Human
    Verified Customer | Posted 11/23/2020
    Human PD‑1 Antibody AF1086
  • Human PD-1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 07/23/2018
    293T cells were infected by lentivirus overexpression of control or PD-1 for 24 and 48 h. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1 um/ml Human PD-1 Antibody (AF1086). A specific band was detected for PD-1 at approximately 43 kDa. This experiment was conducted under reducing conditions
    Human PD‑1 Antibody AF1086
  • Human PD-1 Antibody
    Name: Casey Keyes
    Application: ELISA
    Sample Tested: Human recombinant antibody
    Species: Human
    Verified Customer | Posted 03/07/2018
  • Human PD-1 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Human recombinant test antibody
    Species: Human
    Verified Customer | Posted 09/29/2017
  • Human PD-1 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Species: Human
    Verified Customer | Posted 03/14/2017
    10^6 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.
    Human PD‑1 Antibody AF1086
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Mouse salivary gland (one injected with human T cells and one with no human T cells)
    Species: Human
    Verified Customer | Posted 07/09/2016
    Human PD‑1 Antibody AF1086

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