Recombinant Human PD-1 Fc Chimera Alexa Fluor® 647 Protein

Streptavidin coated beads conjugated to biotinylated anti-human PD‑1 Monoclonal Antibody were stained with the indicated concentrations of Recombinant Human PD‑1 Fc Chimera Alexa Fluor® 647 (Catalog # AFR1086).

2 μg/lane of Recombinant Human PD-1 Fc Chimera Alexa Fluor® 647 Protein (Catalog # AFR1086) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 60-70 kDa and 120-140 kDa, respectively.

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human PD-1 Fc Chimera Alexa Fluor® 647 Protein Summary

• Details of Functionality: Measured by flow cytometry for its ability to bind anti-human PD-1 Monoclonal Antibody conjugated beads.The concentration of Recombinant Human PD-1 Fc Chimera Alexa Fluor® 647 (Catalog # AFR1086) that produces 50% of the binding response is 0.25‑5.00 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived human PD-1 protein
  

  Human PD-1 (Leu25-Gln167) Accession # Q15116.3	IEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: Q15116.3
• N-terminal Sequence: Leu25
• Structure / Form: Disulfide-linked homodimer
  Labeled with Alexa Fluor® 647
  Excitation Wavelength: 650 nm
  Emission Wavelength: 668 nm
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 42.6 kDa (monomer).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 60-70 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Notes

This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human PD-1 Fc Chimera Alexa Fluor® 647 Protein

• CD279 antigen
• CD279
• hPD-1
• PD1
• PD-1
• PD1hPD-l
• PDCD1
• programmed cell death 1
• programmed cell death protein 1
• Protein PD-1
• SLEB2

Background

PD-1 (Programmed Death-1 receptor), also known as CD279, is a receptor expressed on T cells responsible for modulating T cell activation. Like CTLA‑4, PD-1 is classified as an immune checkpoint inhibitory receptor. When PD-1 protein binds to PD-L1, it initiates a negative signaling cascade inhibiting activation of T cells. 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. Normally, PD-1 helps keep T cells from attacking other cells in the body. However, many cancers take advantage of this by expressing high amounts of PD-L1 allowing cancer cells to evade the body's own immune response. Blocking the PD-1:PD-L1 interaction has proven successful in treating many different cancer types. PD-1 protein is type I transmembrane receptor 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 an extracellular region (ECD) with one immunoglobulin-like V‑type domain, a transmembrane domain, and a cytoplasmic region. The mature ECD of human PD-1 shares 61% amino acid sequence identity with mouse PD-1 ECD. PD-1 protein 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).

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.

Bioinformatics

• Uniprot:
  • Q15116.3()