General description
33.47/30.00/31.45/32.10 kDa (mature isoform OA3-323/OA3-293/OA3-305/OA3-312) calculated. ~60-65 kDa (glycosylated) and ~35 kDa (deglycosylated) reported (Parkos, C.A., et al. (1996). J. Cell Biol. 132(3):437-450).
Leukocyte surface antigen CD47 (UniProt Q08722; also known as Antigenic surface determinant protein OA3, CD47, IAP, Integrin-associated protein, Protein MER6) is encoded by the CD47 (also known as MER6) gene (Gene ID 861) in human. Originally defined as a tumor-specific marker for ovarian carcinoma, CD47 a multitransmembrane glycoprotein widely expressed on all hematopoietic cells and most other cell types. CD47 plays a role in αvβ3-mediated cell adhesion and Ca2+ influx, as well as in platelet activation via interaction with thrombospondin-1. In addtion, CD47 modulates peripheral mononuclear (PMN) neutrophil migration across endothelial and epithelial monolayers. The rate of fMLP-stimulated PMN migration is shown to be decreased by CD47 functional blockage upon neutralizaing antibody treatment, which can be partially reversed by PI3K inhibition. Following fMLP stimulation and after PMN transmigration, secondary granules-derived CD47 molecules are shown to accumulate on plasma membrane, resulting in increased cell surface CD47 immunoreactivity. CD47 is produced with a signal peptide sequence (a.a. 1-18) that is removed posttranslationally to yiled the 5-transmembrane (a.a. 142-162, 177-197, 208-228, 236-256, 269-289) protein with its N-terminal end (a.a. 19-141), composed almost entirely of a V-type Ig-like domain (a.a. 19-127), exposed extracellularly and its C-terminal end (a.a. 290-323) at the cytoplasmic side.
Immunogen
CD47 purified from human spleen cells (Liu, Y., et al. (2001). J. Biol. Chem. 276(43):40156-40166).
Epitope: extracellular domain
Application
Research Category
Inflammation & Immunology
This mouse monoclonal Anti-CD47, clone PF3.1 Antibody, Cat. No. MABF958 is validated for use in Flow Cytometry, Immunoprecipitation, and Western Blotting for the detection of CD47.
Immunoprecipitation Analysis: A representative lot immunoprecipitated an increased amount of surface CD47 from human polymorphonuclear (PMN) neutrophils following fMLP stimulation and after transmigration (Parkos, C.A., et al. (1996). J. Cell Biol. 132(3):437-450).
Western Blotting Analysis: A representative lot detected CD47 distribution among human polymorphonuclear (PMN) neutrophil subcellular fractions. Majority of CD47 was found co-localized with CD11b in the CD11b and secondary granules in unstimulated PMN and upregulated plasma membrane localization was seen following fMLP stimulation (Parkos, C.A., et al. (1996). J. Cell Biol. 132(3):437-450).
Biochem/physiol Actions
Clone PF3.1 detects human CD47 by targeting CD47 extracellular domain. In contrast to clone C5/D5 (Cat. No. MABF959), clone PF3.1 is suitable for immunoblotting, but not suitable for neutralizing CD47-dependent cellular signaling, while both clones can stain cell surface CD47 (Liu, Y., et al. (2001). J. Biol. Chem. 276(43):40156-40166).
Physical form
Format: Purified
Protein G purified.
Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Preparation Note
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
Evaluated by Flow Cytometry in human PBMCs.
Flow Cytometry Analysis: 0.2 µL of this antibody detected CD47 surface expression on the gated lymphocytes population among one million human PBMCs.
Other Notes
Concentration: Please refer to lot specific datasheet.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | mouse |
| Quality Segment | 100 |
| conjugate | unconjugated |
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| clone | PF3.1, monoclonal |
| species reactivity | human |
| technique(s) | flow cytometry: suitable |
| isotype | IgG1κ |
| NCBI accession no. | NP_001768 |
| UniProt accession no. | Q08722 |
| shipped in | ambient |
| target post-translational modification | unmodified |
| Gene Information | human ... CD47(961) |
