Recombinant Human PD-L1/B7-H1 His Alexa Fluor® 647 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 His-tag Alexa Fluor® 647 (Catalog # AFR9049).

2 μg/lane of Recombinant Human PD-L1/B7-H1 His-tag Alexa Fluor® 647 Protein (Catalog # AFR9049) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 31-42 kDa.

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human PD-L1/B7-H1 His Alexa Fluor® 647 Protein Summary

• Additional Information: His-tag
• 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 His-tag Alexa Fluor® 647 (Catalog # AFR9049) that produces 50% of the binding response is 0.500‑5.00 ng/mL.
• Source: Human embryonic kidney cell, HEK293-derived human PD-L1/B7-H1 protein
  Phe19-Thr239, with a C-terminal 6-His tag
• Accession #: Q9NZQ7.1
• N-terminal Sequence: Phe19
• Structure / Form: Labeled with Alexa Fluor® 647
  Excitation Wavelength: 650 nm
  Emission Wavelength: 668 nm
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, 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: 26 kDa.
  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: 31-42 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 and NaCl with BSA as a carrier protein.
• Purity: >95%, 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-L1/B7-H1 His Alexa Fluor® 647 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

B7-H1, also known as PD-L1 and CD274, is an approximately 65 kDa transmembrane glycoprotein in the B7 family of immune regulatory molecules (1). Mature human B7-H1 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 (2). Within the ECD, human B7-H1 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 (3). B7-H1 is expressed on inflammatory-activated immune cells including macrophages, T cells, and B cells (4-7), keratinocytes (8, 9), enothelial and intestinal epithelial cells (8, 10), as well as a variety of carcinomas and melanoma (11, 12). B7-H1 binds to T cell B7-1/CD80 and PD-1 (7, 8, 12-15). It suppresses T cell activation and proliferation (5, 8, 14, 16) and induces the apoptosis of activated T cells (11). It plays a role in the development of immune tolerance by promoting T cell anergy (7, 14) and enhancing regulatory T cell development (16). B7-H1 favors the development of anti-inflammatory IL-10 and IL-22 producing dendritic cells (5, 10) and inhibits the development of Th17 cells (16). In cancer, B7-H1 provides resistance to T cell mediated lysis, enhances EMT, and enhances the tumorigenic function of Th22 cells (6, 9, 12, 15).

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

Bioinformatics

• Uniprot:
  • Q9NZQ7.1()