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Multiparameter flow cytometric analysis of CD64 expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV480 Mouse IgG1 Isotype Control (Cat. No. 565652; Left Plot) or BD Horizon BV480 Mouse Anti-Human CD64 antibody (Cat. No. 566129/566178; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter contour plots showing the correlated expression of CD64 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Horizon™ BV480 Mouse Anti-Human CD64
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
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
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.
- BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
- 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 10.1 monoclonal antibody specifically binds to CD64, a 72 kDa type I transmembrane glycoprotein that is a high affinity receptor for human IgG (FcγRI), especially the IgG1 and IgG3 subclasses. CD64 is expressed on monocytes, macrophages, dendritic cells, granulocytes activated with interferon-gamma and early myeloid lineage cells. CD64 associates with a signaling FcRγ homodimer to form the functional high affinity FcγRI complex. CD64 functions in both innate and adaptive immune responses and mediates endocytosis, phagocytosis, antigen presentation, antibody-dependent cellular toxicity, cytokine release and superoxide generation.
The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set. BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.
Development References (5)
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CD64. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:151.
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Dougherty GJ, Selvendran Y, Murdoch S, Palmer DG, Hogg N. The human mononuclear phagocyte high-affinity Fc receptor, FcRI, defined by a monoclonal antibody, 10.1. Eur J Immunol. 1987; 17(10):1453-1459. (Immunogen: Blocking, Flow cytometry, Inhibition). View Reference
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Indik ZK, Hunter S, Huang MM, et al. The high affinity Fc gamma receptor (CD64) induces phagocytosis in the absence of its cytoplasmic domain: the gamma subunit of Fc gamma RIIIA imparts phagocytic function to Fc gamma RI. Exp Hematol. 1994; 22(7):599-606. (Biology). View Reference
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Sun W, O'Shea JJ, Guyre PM. CD64 Workshop Panel Report. In: 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.; 1997:988-990.
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van Vugt MJ, Heijnen AF, Capel PJ, et al. FcR gamma-chain is essential for both surface expression and function of human Fc gamma RI (CD64) in vivo. Blood. 1996; 87(9):3593-3599. (Biology). View Reference
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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.