Description: The monoclonal antibody B6H12 reacts to CD47 also known as integrin-associated protein (IAP), and neurophilin. CD47 is a glycosylated five transmembrane protein with a small alternatively spliced cytoplasmic domain. CD47 is involved in adhesion through interactions with SIRP (signal regulator protein) and is non-covalently associated with beta3 integrins CD51/CD61 and CD41/CD61. Furthermore this interaction can mediate bi-directional signaling to modify neural synaptic activity and regulate the phagocytic activities of macrophages. CD47 is the receptor for thrombospondin. T cell expression of CD47 can mediate activation or apoptosis (in the presence of high levels of thrombospondin). Recently stimulation of CD47 by monoclonal antibody has been shown to induce CD4+CD25- suppressive activity also increasing expression of Foxp3. Expression is found in the majority of hematopoietic cells including T and B cells, monocytes, platelets and erythrocytes (as part of the Rh complex). Expression is also found in non-hematopoietic cells.
This antibody has been reported to have neutralizing activity.
Applications Reported: This B6H12 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This B6H12 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CD47 (integrin-associated protein, IAP) is a ubiquitously expressed cell surface transmembrane glycoprotein interacting with several integrins and regulating their functions. Engagement of CD47 by soluble ligands or counter receptors modulates various signaling pathways, such as activation of heterotrimeric G proteins. Functionally, CD47 is involved in the increase in intracellular calcium concentration that occurs upon cell adhesion to extracellular matrix. CD47 is also a receptor for the C-terminal cell binding domain of thrombospondin, and it may play a role in membrane transport and signal transduction. CD47 has broad tissue distribution, is reduced in expression on Rh erythrocytes. CD47 acts as a ligand for CD172a (signal regulatory protein alpha, SIRP alpha), an immune inhibitory receptor on macrophages; this interaction prevents phagocytosis of CD47-positive cells. Moreover, CD47-CD172a system affects cell migration, B cell adhesion, T cell activation, and is involved in modulation of chondrocyte responses to mechanical signals, and promotes neuronal development, being especially abundant in synapse-rich regions of brain and retina.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: antigen identified by monoclonal antibody 1D8; Antigenic surface determinant protein OA3; CD47; CD47 antigen; CD47 antigen (Rh-related antigen, integrin-associated signal transducer); CD47 glycoprotein; IAP; integrin associated protein; Integrin-associated protein; integrin-associated signal transducer; Leukocyte surface antigen CD47; MER6; OA3; Protein MER6; Rh-related antigen
Gene Aliases: CD47; IAP; MER6; OA3
UniProt ID: (Human) Q08722
Entrez Gene ID: (Human) 961
Molecular Function: G-protein modulator cell adhesion molecule cytokine receptor defense/immunity protein enzyme modulator guanyl-nucleotide exchange factor immunoglobulin receptor superfamily receptor