Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF

When Recombinant Human SIRP alpha/CD172a Fc Chimera (4546-SA) is immobilized at 0.1 µg/mL (100 µL/well), Recombinant Human CD47 Fc Chimera Avi-tag (Catalog # AVI4670) binds with an ED50 of 6-48 ng/mL.

2 μg/lane of Recombinant Human CD47 Fc Chimera Avi-tag (Catalog # AVI4670) 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.

Avi-tag Biotinylated Recombinant Human CD47 Fc protein (Catalog # AVI4670) was immobilized on a Biacore Sensor Chip CM5 via the Avi-tag biotin, and binding to Recombinant Human SIRP alpha His protein (9378-SA) was measured at a concentration range between 6.85 nM and 3.51 µM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=0.134 µM.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human SIRP alpha/CD172a Fc Chimera (Catalog # 4546-SA) is immobilized at 0.1 μg/mL (100 μL/well), the concentration of Recombinant Human CD47 Fc Chimera Avi-tag (Catalog # AVI4670) that produces 50% of the optimal binding response is 6‑48 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human CD47 protein
  

  Human CD47 (Gln19-Pro139) Accession # Q08722	IEGRMD	Human IgG1 (Pro100-Lys330)	Avi-tag
  N-terminus			C-terminus

• Accession #: Q08722
• N-terminal Sequence: No results obtained: Gln19 predicted
• Structure / Form: Disulfide-linked homodimer, Biotinylated via Avi-tag
• 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: 42 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: 60-70 kDa, under reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF

• antigen identified by monoclonal 1D8
• Antigenic surface determinant protein OA3
• CD47 antigen (Rh-related antigen, integrin-associated signal transducer)
• CD47 antigen
• CD47 glycoprotein
• CD47 molecule
• CD47
• IAP
• IAPintegrin associated protein
• Integrin-associated protein
• leukocyte surface antigen CD47
• MER6
• MER6integrin-associated signal transducer
• OA3
• Protein MER6
• Rh-related antigen

Background

CD47, also known as Integrin-Associated Protein (IAP) and OA3, is a 40-60 kDa variably glycosylated atypical member of the immunoglobulin superfamily (1, 2). Human CD47 is an integral membrane protein that consists of a 123 amino acid (aa) extracellular domain (ECD) with a single Ig-like domain, five membrane-spanning regions with short intervening loops, and a 34 aa C-terminal cytoplasmic tail (3). Alternate splicing of human CD47 generates additional isoforms with deletions in the cytoplasmic tail (3). Within the N-terminal ECD, human CD47 shares 63% aa sequence identity with mouse and rat CD47. A portion of the N‑terminal ECD can by shed from smooth muscle cells by MMP-2-mediated proteolysis (4). The ubiquitously expressed CD47 binds to SIRP family members on macrophages, neutrophils, and T cells (5, 6). These interactions prevent macrophage-mediated clearance of healthy CD47-expressing cells and promote immune cell transmigration across the vascular endothelium (5-8). The CD47-SIRP alpha interaction is species specific, and this lack of cross-species interaction has been implicated in xenotransplantation rejection (16). CD47 associates in cis with Fas on T cells and enhances Fas‑mediated apoptosis; its ligation promotes T cell anergy and dampens Th1 immune responses (9-11). CD47 also associates in cis with Integrins  alpha 4 beta 1, alpha V beta 3, alpha 2b beta 3, and  alpha 2 beta 1 which can positively or negatively modulate Integrin‑mediated function (2, 12). In the vasculature, CD47 binding by Thrombospondin-1 inhibits the angiogenic and vasorelaxant effects of nitric oxide (2, 13, 14). On dendritic cells and myeloma cells, CD47 ligation by TSP-1 induces giant cell formation and osteoclast differentiation (15).

1. Sarfati, M. et al. (2009) Curr. Drug Targ. 9:842.
2. Isenberg, J.S. et al. (2008) Arterioscler. Thromb. Vasc. Biol. 28:615.
3. Campbell, I.G. et al. (1992) Cancer Res. 52:5416.
4. Maile, L.A. et al. (2008) Mol. Endocrinol. 22:1226.
5. Oldenborg, P.-A. et al. (2000) Science 288:2051.
6. Liu, Y. et al. (2002) J. Biol. Chem. 277:10028.
7. Stefanidakis, M. et al. (2008) Blood 112:1280.
8. de Vries, H.E. et al. (2002) J. Immunol. 168:5832.
9. Manna, P.P. et al. (2005) J. Biol. Chem. 280:29637.
10. Avice, M.-N. et al. (2001) J. Immunol. 167:2459.
11. Bouguermouh, S. et al. (2008) J. Immunol. 180:8073.
12. Barazi, H.O. et al. (2002) J. Biol. Chem. 277:42859.
13. Isenberg, J.S. et al. (2009) Nitric Oxide 21:52.
14. Isenberg, J.S. et al. (2009) Matrix Biol. 28:110.
15. Kukreja, A. et al. (2009) Blood 114:3413.
16. Wang H. et al. (2007) Blood 109:836.

Bioinformatics

• Uniprot:
  • Q08722()