Recombinant Human PD-L1/B7-H1 Fc Alexa Fluor® 488 Protein

Streptavidin coated beads conjugated to biotinylated anti-human PD-L1/B7-H1 Monoclonal Antibody were stained with the indicated concentrations of Recombinant Human PD-L1/B7-H1 Fc Chimera Alexa Fluor® 488 (Catalog # AFG156).

2 μg/lane of Recombinant Human PD-L1/B7-H1 Fc Chimera Alexa Fluor® 488 Protein (Catalog # AFG156) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70-75 kDa, and 140-150 kDa, respectively.

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human PD-L1/B7-H1 Fc Alexa Fluor® 488 Protein Summary

• Additional Information: Fc Chimera
• Details of Functionality: Measured by flow cytometry for its ability to bind anti-human PD-L1/B7-H1 Monoclonal Antibody conjugated beads. The concentration of Recombinant Human PD-L1/B7-H1 Fc Chimera Alexa Fluor® 488 (Catalog # AFG156) that produces 50% of the binding response is 15.0‑150 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived human PD-L1/B7-H1 protein
  

  Human PD-L1 (Phe19-Thr239) Accession # Q9NZQ7.1	DIEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: Q9NZQ7.1
• N-terminal Sequence: Phe19
• Structure / Form: Disulfide-linked homodimer
  Labeled with Alexa Fluor® 488 via amines
  Excitation Wavelength: 488 nm
  Emission Wavelength: 515-545 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: 52 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: 70-75 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
• Buffer: Supplied as a 0.2 μm filtered solution in PBS and NaCl 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 produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human PD-L1/B7-H1 Fc Alexa Fluor® 488 Protein

• Avelumab
• B7-H
• B7H1
• B7-H1
• B7H1PDCD1L1
• CD274 antigenMGC142294
• CD274 molecule
• CD274
• PDCD1L1
• PDCD1LG1
• PDCD1LG1MGC142296
• PDL1
• PD-L1
• PD-L1B7 homolog 1
• PDL1PDCD1 ligand 1
• programmed cell death 1 ligand 1
• Programmed death ligand 1

Background

PD-L1, also known as B7-H1, PDL1, is one of the ligands for PD-1 and plays a critical role in the regulation of T cell immunity (1-6). The PD-1:PD-L1 interaction initiates a negative signaling cascade in T cells leading to inhibition of T cell activation (2, 5, 7, 8). PD-L1 provides a molecular stop signal to the adaptive immune system helping to distinguish between self and foreign antigens. PD-L1 also plays a role in the development of immune tolerance by promoting T cell anergy (1, 5) and enhancing regulatory T cell development (8). In addition, PD-L1 favors the development of anti-inflammatory IL-10 and IL-22 producing dendritic cells (7, 9) and inhibits the development of Th17 cells (8). Many cancers exhibit upregulated PD-L1 protein expression, and several cancers with high levels of PD-L1 have been associated with increased tumor aggressiveness and poor prognosis. Using new therapeutics that block the PD-L1:PD-1 interaction has proven successful in the clinic for many cancer types and has sparked great interest in the field of cancer immunotherapy.

The PD-L1 protein is an approximately 65 kDa transmembrane glycoprotein belonging to the B7 family of immune regulatory molecules (10). Mature human PD-L1 protein consists of a 220 amino acid (aa) extracellular domain (ECD) with two immunoglobulin-like domains, a 21 aa transmembrane segment, and a 31 aa cytoplasmic domain (11). Within the ECD, human PD-L1 shares 73% and 74% aa sequence identity with mouse and rat B7-H1, respectively. Alternative splicing generates additional isoforms that either lack the first Ig-like domain or are truncated within the second Ig-like domain (12). PD-L1 is expressed on inflammatory-activated immune cells including macrophages, T cells, and B cells (10, 13, 14, 16) keratinocytes (9, 11), endothelial and intestinal epithelial cells (2, 9), as well as a variety of carcinomas and melanoma (12, 16).

1. Tsushima, F. et al. (2007) Blood 110:180.
2. Mazanet, M.M. and C.C.W. Hughes (2002) J. Immunol. 169:3581.
3. Azuma, T. et al. (2008) Blood 111:3635.
4. Butte, M.J. et al. (2008) Mol. Immunol. 45:3567.
5. Park, J.-J. et al. (2010) Blood 116:1291.
6. Ritprajak, P. et al. (2010) J. Immunol. 184:4918.
7. Chen, L. et al. (2007) J. Immunol. 178:6634.
8. Herold, M. et al. (2015) J. Immunol. 195:3584.
9. Scandiuzzi, L. et al. (2014) Cell Rep. 6:625.
10. Ceeraz, S. et al. (2013) Trends Immunol. 34:556.
11. Dong, H. et al. (1999) Nat. Med. 5:1365.
12. Frigola, X. et al. (2011) Clin. Cancer Res. 17:1915.
13. Tamura, H. et al. (2001) Blood 97:1809.
14. Kuang, D.-M. et al. (2014) J. Clin. Invest. 124:4657.
15. Cao, Y. et al. (2010) Cancer Res. 71:1235.
16. Dong, H. et al. (2002) Nat. Med. 8:793.

Bioinformatics

• Uniprot:
  • Q9NZQ7.1()