Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Western Blot, ELISA, Flow Cytometry, CyTOF-ready

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2335A
View all PD-1 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line, NS0-derived human PD-1
Met1-Gln167
Accession # Q15116

Specificity

Detects human PD-1 in direct ELISAs.

Clonality

Monoclonal

Host

Rabbit

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 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 antibody in Human PBMCs antibody by Flow Cytometry.

Detection of PD‑1 in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) either (A) treated with 5 ug/mL PHA for 5 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 Antibody in ELISA Standard Curve.

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).

Applications for Human PD‑1 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

ELISA

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.

Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMC treated with PHA

Western Blot

0.5 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with human PD-1

Reviewed Applications

Read 1 review rated 4 using MAB10863 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: PD-1

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).

References

  1. Ishida, Y. et al. (1992) EMBO J. 11:3887.
  2. Sharpe, A.H. and G. J. Freeman (2002) Nat. Rev. Immunol. 2:116.
  3. Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
  4. Nishimura, H. and T. Honjo (2001) Trends Immunol. 22:265.
  5. Watanabe, N et al. (2003) Nat. Immunol. 4:670.
  6. Zhang, X. et al. (2004) Immunity 20:337.
  7. Lázár-Molnár, E. et al. (2008) Proc. Natl. Acad. Sci. USA 105:10483.
  8. Nishimura, H et al. (1996) Int. Immunol. 8:773.
  9. Keir, M.E. et al. (2008) Annu. Rev. Immunol. 26:677.
  10. Butte, M.J. et al. (2007) Immunity 27:111.
  11. Okazaki, T. et al. (2013) Nat. Immunol. 14:1212.
  12. Iwai, Y. et al. (2002) Proc. Natl. Acad. Sci. USA 99: 12293.
  13. Nogrady, B. (2014) Nature 513:S10.
  14. Swaika, A. et al. (2015) Mol. Immunol. 67: 4

Long Name

Programmed Death-1

Alternate Names

CD279, PD1, PDCD1, SLEB2

Entrez Gene IDs

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

Gene Symbol

PDCD1

UniProt

Q15116

Additional PD-1 Products

Product Documents for Human PD‑1 Antibody

Certificate of Analysis

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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

Customer Reviews for Human PD‑1 Antibody (1)

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  • Human PD-1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 08/28/2023
    A Western blot analysis was performed using lysates from the HEK293T human embryonic kidney cell line that had been transfected with human PD-1. A PVDF membrane was utilized and probed with the antibody at a concentration of 0.5 µg/mL. The bands corresponding to PD-1 were observed at around 70 kDa. The experimental conditions involved conducting the assay under reducing conditions.
    Human PD‑1 Antibody MAB10863

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Protocols

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