United Kingdom (UK)
Recombinant monoclonal antibody to CD47. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the phage display antibody C47B227.
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: Three de novo phage libraries displaying Fab variants on phage coat protein pIX were panned against biotinylated human CD47.
Specificity: The antibody is specific for CD47. The antibody binds a conformational epitope composed of residues in the C’C’’ loop (L54) and B-strand C*’ (K56 and T58) regions.
Application Notes: The structure of the Fab fragment in complex with CD47 was determined. The binding affinity of the antibody to human and cyno CD47 were measured (Kd= 1nM and 0,5 nM respectively). The antibody inhibited CD47 from interacting with SIRP alpha (US10864268).
Antibody first published in: PMID:
Western Blot using anti-CD47 antibody C47B227 (Ab03167). Human placenta(A) (0.03µg/ml) and human prostate(B) (0.01ug/ml) tissue lysates lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of C47B227 (Ab03167-23.0) 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 (Ab03167) C47B227 Immunofluorescence analysis of paraformaldehyde fixed Jurkat cells immobilized on Shi-fix™ plus cover-slips stained with the chimeric rabbit IgG version of C47B227 (Ab03167-23.0) (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 Ab03167-23.0, 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 C47B227 (Ab03167). Paraformaldehyde fixed Jurkat cells were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of C47B227 (Ab03167-23.0, 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.