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Multiparameter flow cytometric analysis of CD47 expression on human peripheral blood leucocyte populations. Human peripheral blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were stained with either BD Pharmingen™ PE Mouse IgG1, κ Isotype Control (Cat. No. 556650; Left Plot) or BD Pharmingen™ PE Mouse Anti-Human CD47 antibody (Cat. No. 568089/568090; Right Plot). A bivariate pseudocolor density plot showing the correlated expression of CD47 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ PE Mouse Anti-Human CD47
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BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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Companion Products
The CC2C6 monoclonal antibody specifically recognizes CD47 which is also known as Integrin-associated protein (IAP), Rh-related antigen, Ovarian carcinoma antigen (OA3), MER6, or neutrophilin. CD47 is a 42-52 kDa N-linked glycan protein which has an extracellular N-terminal IgV-like domain followed by 5 transmembrane segments and a short C-terminal cytoplasmic domain. CD47 is expressed on all hematopoietic cells, including leucocytes, platelets, and erythrocytes except for Rh-negative erythrocytes. It is also expressed on epithelial cells, endothelial cells, fibroblasts, and many tumor cell lines. CD47 binds to and activates the inhibitory Signal regulatory protein α (SIRPα) expressed on phagocytic cells. This trans interaction inhibits phagocytosis and protects CD47-positive cells, including "self" blood cells like erythrocytes, from phagocytosis. CD47 binds to thrombospondin-1 which regulates cell adhesion and migration and plays a role in the regulation of inflammation and angiogenesis. CD47 can also interact with integrins which can affect various cellular functions including cellular adhesion, spreading, and migration. The CC2C6 antibody can reportedly inhibit the interaction between CD47 and SIRPα.
Development References (4)
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Lindberg FP, Gresham HD, Schwarz E, Brown EJ. Molecular cloning of integrin-associated protein: an immunoglobulin family member with multiple membrane-spanning domains implicated in alpha v beta 3-dependent ligand binding. J Cell Biol. 1993; 123(2):485-496. (Biology). View Reference
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Lindberg FP, Lublin DM, Telen MJ, et al. Rh-related antigen CD47 is the signal-transducer integrin-associated protein. J Biol Chem. 1994; 269(3):1567-1570. (Biology). View Reference
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Seiffert M, Cant C, Chen Z, et al. Human signal-regulatory protein is expressed on normal, but not on subsets of leukemic myeloid cells and mediates cellular adhesion involving its counterreceptor CD47. Blood. 1999; 94(11):3633-3643. (Immunogen: Blocking, Flow cytometry). View Reference
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Zola H, Swart B, Nicholson I, Voss E. CD47. In: Zola H. Leukocyte and Stromal Cell Molecules : the CD Markers. Hoboken, N.J.: Wiley-Liss; 2007:119-120.
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