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Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF

R&D Systems, part of Bio-Techne | Catalog # AVI4670

R&D Systems, part of Bio-Techne
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AVI4670-050

Key Product Details

Learn more about Avi-tag Biotinylated Proteins

Source

CHO

Accession #

Structure / Form

Disulfide-linked homodimer, Biotinylated via Avi-tag

Conjugate

Biotin

Applications

Bioactivity

Product Specifications

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

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

No results obtained: Gln19 predicted

Predicted Molecular Mass

42 kDa

SDS-PAGE

60-70 kDa, under reducing conditions

Activity

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.

Scientific Data Images for Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF

Recombinant Human CD47 Fc Chimera Avi-tag Protein Bioactivity

Recombinant Human CD47 Fc Chimera Avi-tag Protein Bioactivity

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.
Recombinant Human CD47 Fc Chimera Avi-tag Protein SDS-PAGE

Recombinant Human CD47 Fc Chimera Avi-tag Protein SDS-PAGE

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.
Surface plasmon resonance sensorgram of Avi-tag biotinylated human CD47 protein binding to human SIRP alpha

Binding of Human SIRP alpha to CD47 Fc Avi-tag Protein by surface plasmon resonance (SPR).

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.

Formulation, Preparation and Storage

AVI4670
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: CD47

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  alpha4 beta1, alphaV beta3, alpha2b beta3, and  alpha2 beta1 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).

References

  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.

Alternate Names

CD47, IAP, MER6, OA3

Entrez Gene IDs

961 (Human); 16423 (Mouse); 397042 (Porcine); 478552 (Canine); 102139846 (Cynomolgus Monkey)

Gene Symbol

CD47

UniProt

Additional CD47 Products

Product Documents for Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human CD47 Fc Chimera Avi-tag Protein, CF

For research use only

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