Recombinant Human B7-H2 Fc Chimera Alexa Fluor® 647 Protein

Streptavidin coated beads conjugated to Human B7-H2 Biotinylated Antibody were stained with the indicated concentrations of Recombinant Human B7‑H2 Fc Chimera Alexa Fluor® 647 (Catalog # AFR165).

2 μg/lane of Recombinant Human B7‑H2 Fc Chimera Alexa Fluor® 647 Protein (Catalog # AFR165) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 80-90 kDa and 160-180 kDa, respectively.

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human B7-H2 Fc Chimera Alexa Fluor® 647 Protein Summary

• Details of Functionality: Measured by flow cytometry for its ability to bind anti-human B7-H2 Antibody conjugated beads. The concentration of Recombinant Human B7‑H2 Fc Chimera Alexa Fluor® 647 (Catalog # AFR165) that produces 50% of the binding response is 2.50‑25.0 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived human B7-H2 protein
  

  Human B7-H2 (Asp19-Ser258) Accession # O75144.2	DIEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: O75144.2
• N-terminal Sequence: Asp19
• Structure / Form: Disulfide-linked homodimer
  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: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 53.4 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: 80-90 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: >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 B7-H2 Fc Chimera Alexa Fluor® 647 Protein

• B7H2
• B7-H2
• B7H2B7 homolog 2
• B7-H2B7-related protein 1
• B7RP-1
• B7RP1B7-like protein Gl50
• B7RP-1LICOS
• CD275 antigen
• CD275
• GL50
• GL50transmembrane protein B7-H2 ICOS ligand
• ICOSL
• ICOS-L
• ICOSLG
• inducible T-cell co-stimulator ligand
• KIAA0653ICOS ligand

Background

B7-H2, also known as B7-related protein (B7RP1), ICOS Ligand, and CD275, is an approximately 60 kDa transmembrane glycoprotein in the B7 family of immune regulatory molecules (1). Mature human B7-H2 consists of a 238 amino acid (aa) extracellular domain (ECD) with two immunoglobulin-like domains, a 21 aa transmembrane segment, and a 25 aa cytoplasmic domain (2, 3). Within the ECD, human B7-H2 shares 50% and 54% aa sequence identity with mouse and rat B7‑H2, respectively. Alternative splicing generates a long isoform that carries a 10 aa substitution for the C-terminal 3 residues. B7-H2 is expressed on antigen presenting cells such as B cells, macrophages, monocytes, and dendritic cells (2-6). It binds to ICOS on activated T cells, leading to both positive and negative effects on immune responses including its own down-regulation (2, 4, 7). Mouse and human B7-H2 exhibit cross-species binding to ICOS (3, 6). The B7-H2 interaction with ICOS is costimulatory for T cell proliferation as well as the development of B cells, plasma cells, follicular helper T cells (T^FH) and germinal centers (2-4, 8, 9). In human but not in mouse, B7-H2 additionally binds to CD28 and CTLA4, and its interaction with CD28 can costimulate both human and mouse naïve T cells and regulatory T cells (Treg) (6). B7-H2 contributes to the development of allergic asthma by enhancing Th2 biased immune responses, limiting Th17 responses, and promoting eosinophilic infiltration into the lung (8, 10, 11). Its activation of ICOS on Treg limits pulmonary inflammation and airway hyperresponsiveness, promotes the development of inhalational tolerance, and impairs anti-tumor immunity (5, 12, 13). In contrast, its ligation of ICOS on T^FH cells can increase the severity of autoimmune symptoms (9). A soluble form of human B7-H2 is elevated in the circulation of patients with active systemic lupus erythematosus (14). In the thyroid, B7-H2 is up-regulated on thyrocytes during inflammation and promotes their proliferation and production of thryoid hormones (15).

1. Bour-Jordan, H. et al. (2011) Immunol. Rev. 241:180.
2. Wang, S. et al. (2000) Blood 96:2808.
3. Yoshinaga, S.K. et al. (2000) Int. Immunol. 12:1439.
4. Yoshinaga, S.K. et al. (1999) Nature 402:827.
5. Faget, J. et al. (2012) Cancer Res. 72:6130.
6. Yao, S. et al. (2011) Immunity 32:729.
7. Watanabe, M. et al. (2008) J. Immunol. 180:5222.
8. Wong, S.-C. et al. (2003) Blood 102:1381.
9. Hu, Y.-L. et al. (2009) J. Immunol. 182:1421.
10. Kadkhoda, K. et al. (2010) J. Immunol. 184:3780.
11. Kadkhoda, K. et al. (2011) Int. Immunol. 23:239.
12. Gajewska, B.U. et al. (2005) J. Immunol. 174:3000.
13. Akbari, O. et al. (2002) Nat. Med. 8:1024.
14. Her, M. et al. (2009) Lupus 18:501.
15. Wang, F. et al. (2012) J. Clin. Immunol. 32:1253.

Publications for B7-H2/ICOSLG (AFR165)(1)

Publication using AFR165

• Majocchi, S;Lloveras, P;Nouveau, L;Legrand, M;Viandier, A;Malinge, P;Charreton, M;Raymond, C;Pace, E;Millard, B;Svensson, L;Kelpšas, V;Anceriz, N;Salgado-Pires, S;Daubeuf, B;Magistrelli, G;Gueneau, F;Moine, V;Masternak, K;Shang, L;Fischer, N;Ferlin, W; NI-3201 Is a Bispecific Antibody Mediating PD-L1-Dependent CD28 Co-stimulation on T Cells for Enhanced Tumor Control Cancer Immunology Research 2025-01-06 [PMID: 39760515] (Flow Cytometry, Human)

Bioinformatics

• Uniprot:
  • O75144.2()