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Multiparameter flow cytometric analysis using BD OptiBuild™ BV650 Mouse Anti-Human CD64 antibody (Cat. No. 745316) on human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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).
The MD22 monoclonal antibody specifically recognizes CD64 which is also known as Fc-gamma receptor I (FcgR1) or FcγRI. CD64 is a ~72 kDa type I transmembrane glycoprotein that is encoded by FCGR1A (Fc fragment of IgG receptor Ia). CD64 functions as a high affinity receptor for human IgG. The CD64 antigen is expressed on monocytes, macrophages, at low levels on polymorphonuclear neutrophils (PMNs), and on a subpopulation of circulating dendritic cells. The CD64 antigen is an early granulomonocytic lineage marker on CD34+ hematopoietic progenitors. Expression of the CD64 antigen increases transiently in cases of sepsis. The CD64 antigen functions in both innate and adaptive immune responses, and mediates endocytosis, phagocytosis, antigen presentation, antibody-dependent cellular cytotoxicity, cytokine release, and superoxide generation. The CD64 antigen associates with the signal-transducing γ-chain homodimer of Fc receptors to form the functional high affinity FcγRI complex. Ligation of the CD64 antigen leads to the activation of the protein tyrosine kinases, hck and lyn.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
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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.