Recombinant Human B7-H4 Fc Chimera Alexa Fluor® 647 Protein

Streptavidin coated beads conjugated to biotinylated Human B7‑H4 were stained with the indicated concentrations of Recombinant Human B7‑H4 Fc Chimera Alexa Fluor® 647 Protein (Catalog# AFR8870).

2 μg/lane of Recombinant Human B7‑H4 Fc Chimera Alexa Fluor® 647 Protein (Catalog # AFR8870) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 78-90 kDa and 156-180 kDa, respectively.

• Applications: Bioactivity

Recombinant Human B7-H4 Fc Chimera Alexa Fluor® 647 Protein Summary

• Details of Functionality: Measured by flow cytometry for its ability to bind anti-human B7-H4 Antibody conjugated beads. The concentration of Recombinant Human B7-H4 Fc Chimera Alexa Fluor® 647 (Catalog # AFR8870) that produces 50% of the binding response is 7.50-75.0 ng/mL.
• Source: Human embryonic kidney cell, HEK293-derived human B7-H4 protein
  

  Human B7-H4 (Phe29-Ala258) Accession # Q7Z7D3.1	IEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: Q7Z7D3.1
• N-terminal Sequence: Phe29
• 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: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 52 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: 78-90 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 produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human B7-H4 Fc Chimera Alexa Fluor® 647 Protein

• B7h.5
• B7H4
• B7-H4
• B7H4T-cell costimulatory molecule B7x
• B7S1
• B7S1VCTN1
• B7x
• B7XPRO1291
• FLJ22418
• Immune costimulatory protein B7-H4
• Protein B7S1
• T cell costimulatory molecule B7x
• V-set domain containing T cell activation inhibitor 1
• V-set domain-containing T-cell activation inhibitor 1
• Vtcn1

Background

B7-H4, also known as B7x and B7S1, is a 50-80 kDa glycosylated member of the B7 family of immunomodulatory proteins (1, 2). Mature human B7-H4 consists of a 235 amino acid (aa) extracellular domain (ECD) with one Ig-like V-set domain and one Ig-like C2-set domain, a 21 aa transmembrane segment, and a 2 aa cytoplasmic tail (3-5). Within the ECD, human B7-H4 shares 90% aa sequence identity with mouse and rat B7-H4. It shares 22% - 28% aa sequence identity with human B7-1, B7-2, B7-H1, B7-H2, B7-H3, and PD-L2. Alternate splicing of human B7-H4 generates an additional isoform that lacks the first Ig-like domain. B7-H4 is expressed on the surface of activated lymphocytes, macrophages, monocytes, dendritic cells, epithelial cells, and bone marrow-derived mesenchymal stem cells (4-8). Its binding to activated T cells dampens T cell responses and induces cell cycle arrest in the T cell (3-5). Reverse signaling can induce either cell cycle arrest or apoptosis in the B7-H4 expressing cell (9, 10). B7-H4 is up-regulated in several carcinomas in correlation with tumor progression and metastasis (2, 7, 11, 12). A soluble form of B7-H4 is elevated in the serum of ovarian cancer, renal cell carcinoma, and rheumatoid arthritis patients, also in correlation with advanced disease status (13-15). Soluble B7-H4 functions as a decoy molecule that blocks the inhibitory influence of B7-H4 on immune activation (15). Despite evidence for the involvement of B7-H4 in immune regulation, mice deficient in its expression do not show significant immune deficiencies, suggesting compensation by other molecules in vivo (16).

1. Yi, K.H. and L. Chen (2009) Immunol. Rev. 229:145.
2. Salceda, S. et al. (2005) Exp. Cell Res. 306:128.
3. Zang, X. et al. (2003) Proc. Natl. Acad. Sci. 100:10388.
4. Prasad, V.R. et al. (2003) Immunity 18:863.
5. Sica, G.L. et al. (2003) Immunity 18:849.
6. Kryczek, I. et al. (2006) J. Exp. Med. 203:871.
7. Tringler, B. et al. (2005) Clin. Cancer Res. 11:1842.
8. Xue, Q. et al. (2010) Stem Cells Dev. 19:27.
9. Song, H. et al. (2008) Cancer Lett. 266:227.
10. Park, G.B. et al. (2009) Immunology 128:360.
11. Zang, X. et al. (2007) Proc. Natl. Acad. Sci. 104:19458.
12. Krambeck, A.E. et al. (2006) Proc. Natl. Acad. Sci. 103:10391.
13. Simon, I. et al. (2006) Cancer Res. 66:1570.
14. Thompson, R.H. et al. (2008) Cancer Res. 68:6054.
15. Azuma, T. et al. (2009) PloS Med. 6:e1000166.
16. Suh, W.-K., et al. (2006) Mol. Cell. Biol. 26:6403.

Bioinformatics

• Uniprot:
  • Q7Z7D3.1()