Human PD-1 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB10863-100
MAB10863-SP
Human PD-1 Antibody in Western Blot
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Human PD-1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human PD-1 in direct ELISAs.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2335A
Purification
Protein A or G purified from cell culture supernatant
Immunogen
Mouse myeloma cell line, NS0-derived 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

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
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

Western Blot Detection  of Human PD‑1 by Western Blot. View Larger

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.

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

Detection of PD‑1 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either (A) treated with 5 ng/mL PHA for 2 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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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 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
Entrez Gene IDs
5133 (Human); 18566 (Mouse); 301626 (Rat); 486213 (Canine); 102123659 (Cynomolgus Monkey)
Alternate Names
CD279 antigen; CD279; hPD-1; PD1; PD-1; PD1hPD-l; PDCD1; programmed cell death 1; programmed cell death protein 1; Protein PD-1; SLEB2

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