United Kingdom (UK)
Recombinant monoclonal antibody to CD47. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma B6H12.
UniProt Accession Number of Target Protein: Q08722
Alternative Name(s) of Target: IAP; MER6; OV-3; Leukocyte surface antigen CD47; Antigenic surface determinant protein OA3; Integrin-associated protein; Protein MER6; ovarian cancer marker
Immunogen: The original antibody was generated by immunizing BALB/c mice with purified placental RGD-binding proteins (PRBP).
Specificity: This antibody binds the extracellular domain of the human CD47, which is broadly distributed on normal adult tissues, as well as ovarian tumors, and is especially abundant in some epithelia and the brain. IAP or integrin assocaited protein has a role in both cell adhesion by acting as an adhesion receptor for THBS1 on platelets, and in the modulation of integrins. It further plays an important role in memory formation and synaptic plasticity in the hippocampus. It also acts as a receptor for SIRPA. Binding to SIRPA prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells.
Application Notes: This antibody reacts with placental RGD-binding proteins and human neutrophils in an ELISA (PMID: 2785522). This antibody binds CD47 expressed on human neutrophils in a flow cytometric assay. This antibody immunoprecipitated vitronectin receptor (VnR) from detergent-solubilized placental membrane preparations (PMID: 2785522). It is a function-blocking antibody that inhibits CD47 interactions with TSP1, SIRPα, some integrins, and EGF receptor (PMID: 19636412; 10572074; 19004835; 26840086). This antibody directly induces killing of tumor cells by NK cells, B cell migratiom, and regulatory T cell differentiation (PMID: 18497546; 10891456; 16951312). B6H12 enhanced antibody-dependent cellular phagocytosis (ADCP) of human acute myeloid leukemia (AML) cells in vitro by human and mouse macrophages (PMID: 19632179). THis antibody also inhibited AML tumor growth in NOD/SCID/IL2Rγ− (NSG) mice by blocking this “don’t eat me” signal (PMID: 19632179). B6H12 was used in several preclinical studies to explore combining CD47 blockade with therapeutic antibodies that target specific cancers. This antibody in combination with CD20 antibody rituximab, to promote phagocytosis in vitro and eradicated non-Hodgkin lymphoma in NSG mice (PMID: 20813259).
Antibody first published in:
Gresham et al. A novel member of the integrin receptor family mediates Arg-Gly-Asp-stimulated neutrophil phagocytosis. J Cell Biol. 1989 May;108(5):1935-43. PMID:2785522
Note on publication:
Describes the generation of this antibody and its use in determining the role of CD47 in Arg-Gly-Asp-stimulated neutrophil phagocytosis.
Western Blot using Anti-CD47 antibody B6H12 (Ab03105). Human PBM cells lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of B6H12 (Ab03105-23.0) at 0.3µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of Jurkat cells with Anti-CD47 (Ab03105) B6H12 Immunofluorescence analysis of paraformaldehyde fixed Jurkat cells immobilized on Shi-fix™ plus cover-slips stained with the chimeric rabbit IgG version of B6H12 (Ab03105-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab03105-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using the Anti-CD47 antibody B6H12 (Ab03105). Paraformaldehyde fixed Jurkat cells were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of B6H12 (Ab03105-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.