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Flow cytometric analysis of CD47 expression on human peripheral blood lymphocytes. Whole blood was treated with BD PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were then stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD47 antibody (Cat. No. 563760; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV421 Mouse Anti-Human CD47
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Brilliant Violet™ 421 is a trademark of Sirigen.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The B6H12 monoclonal antibody specifically binds to CD47, a 42-52 kDa N-linked glycan protein. CD47 antigen, also known as integrin-associated protein (IAP), is expressed on all hematopoietic cells, including leukocytes, platelets and erythrocytes. It is also expressed on epithelial cells, endothelial cells, fibroblasts and many tumor cell lines. CD47 may play a role as a signal transducer in the regulation of cation fluxes across cell membranes and in the chemotactic and adhesive interactions of leukocytes with endothelial cells. The B6H12 antibody is capable of blocking phagocytosis of microparticles by peripheral blood granulocytes. It has also been reported to induce proliferation of CD3-activated T cells.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.
Development References (7)
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Brown E, Hooper L, Ho T, Gresham H. Integrin-associated protein: a 50-kD plasma membrane antigen physically and functionally associated with integrins. J Cell Biol. 1990; 111(6):2785-2794. (Clone-specific: Blocking, ELISA, Immunoaffinity chromatography, Immunoprecipitation, Western blot). View Reference
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Gresham HD, Goodwin JL, Allen PM, Anderson DC, Brown EJ. A novel member of the integrin receptor family mediates Arg-Gly-Asp-stimulated neutrophil phagocytosis. J Cell Biol. 1989; 108(5):1935-1943. (Immunogen: ELISA, Flow cytometry, Functional assay, Immunofluorescence, Inhibition). View Reference
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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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. (Clone-specific: Flow cytometry, Immunoprecipitation). 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. (Clone-specific: Immunoaffinity chromatography). View Reference
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.