HEK293 human embryonic kidney cell line transfected with either (A) mouse CD47 or (B) irrelevant protein, and eGFP was stained with Rat Anti-Mouse CD47 Monoclonal Antibody (Catalog # MAB18661) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). Quadrant markers were set based on isotype control antibody (Catalog # MAB005, data not shown). View our protocol for Staining Membrane-associated Proteins.

CD47 Binding to SIRP alpha/CD172a Blocked By Mouse CD47 Antibody.

In a functional ELISA, 0.08-0.8 µg/mL of this antibody (green line) will block 50% of the binding of 0.25 µg/mL of Recombinant Mouse CD47 Fc Chimera (orange line, Catalog # 1866-CD) to immobilized Recombinant Mouse SIRPa/CD172a Fc Chimera (Catalog # 7154-SA) coated at 1 µg/mL (100 µL/well). At 5 µg/mL, this antibody will block >90% of the binding.

Applications for Mouse CD47 Antibody

Application

Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.08-0.8 µg/mL of this antibody will block 50% of the binding of 0.25 µg/mL of Recombinant Mouse CD47 Fc Chimera (Catalog # 1866-CD) to immobilized Recombinant Mouse SIRP alpha/CD172a Fc Chimera (Catalog # 7154-SA) coated at 1 µg/mL (100 µL/well). At 5 µg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 Human Cell Line Transfected with Mouse CD47 and eGFP

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 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). Mouse CD47 is an integral membrane protein that consists of a 122 amino acid (aa) extracellular domain (ECD) with a single Ig‑like domain, five membrane-spanning regions with short intervening loops, and a 16 aa C‑terminal cytoplasmic tail (3). Alternate splicing of mouse CD47 generates an additional isoform with an insertion of 21 aa following the Ig‑like domain (3). Within the N‑terminal ECD, mouse CD47 shares 63% and 84% aa sequence identity with human and rat CD47, respectively. 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

Sarfati, M. et al. (2009) Curr. Drug Targ. 9:842.
Isenberg, J.S. et al. (2008) Arterioscler. Thromb. Vasc. Biol. 28:615.
Lindberg, F.P. et al. (1993) J. Cell Biol. 123:485.
Maile, L.A. et al. (2008) Mol. Endocrinol. 22:1226.
Oldenborg, P.-A. et al. (2000) Science 288:2051.
Liu, Y. et al. (2002) J. Biol. Chem. 277:10028.
Stefanidakis, M. et al. (2008) Blood 112:1280.
de Vries, H.E. et al. (2002) J. Immunol. 168:5832.
Manna, P.P. et al. (2005) J. Biol. Chem. 280:29637.
Avice, M.-N. et al. (2001) J. Immunol. 167:2459.
Bouguermouh, S. et al. (2008) J. Immunol. 180:8073.
Barazi, H.O. et al. (2002) J. Biol. Chem. 277:42859.
Isenberg, J.S. et al. (2009) Nitric Oxide 21:52.
Isenberg, J.S. et al. (2009) Matrix Biol. 28:110.
Kukreja, A. et al. (2009) Blood 114:3413.
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

NP_034711

Additional CD47 Products

Product Documents for Mouse CD47 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Mouse CD47 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Mouse CD47 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB18661

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Mouse CD47 Antibody

Detection of CD47 in HEK293 Human Cell Line Transfected with Mouse CD47 and eGFP by Flow Cytometry.