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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 EBVCS-5 antibody specifically binds to human CD23, the low affinity receptor for human IgE (Fc epsilon RII or FcεRII), which is also known as C-type lectin domain family 4 member J (CLEC4J). CD23 is expressed as a ~45 kDa type II membrane glycoprotein that is present at low density on most normal B lymphocytes and at higher levels on activated B lymphocytes, Epstein-Barr virus (EBV)-transformed lymphoblasts, chronic lymphocytic leukemia (CLL) cells of B-lymphocyte origin, and tonsillar B lymphocytes. CD23 expression is induced on B cells by interleukin-4 (IL-4) and is lost after isotype switching to IgA, IgG, or IgE. The CD23 antigen is not present on immature bone marrow B lymphocytes or on T lymphocytes. CD23 may also be differentially expressed on monocytes, macrophages, eosinophils, platelets, dendritic cells, and Langerhans cells. CD23 can mediate IgE-dependent cytotoxicity and phagocytosis by macrophages and eosinophils. Soluble CD23 (sCD23) can be released by CD23-positive cells as a result of proteolytic cleavage of membrane CD23. Larger fragments of sCD23 (eg, 25-37 kDa) retain their IgE-binding capacity whereas smaller fragments (ie, ≤ 12 kDa) do not. Soluble CD23 may have immunoregulatory effects on the growth and differentiation of B cells and other cell types.
The antibody was conjugated to BD Horizon™ BV711 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 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 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.
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