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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 5H10 monoclonal antibody specifically binds to mouse CD209a, or CIRE, which is the most closely related homolog of human DC-SIGN. In the mouse, several homologs of human CD209 (DC-SIGN) have been described. CD209a is a 238-amino acid type-II transmembrane C-type lectin, with an extracellular carbohydrate recognition domain. It can bind to mannosylated ligands in a Ca++ dependent manner. CD209a is expressed by some conventional CD4+ and CD4-CD8- dendritic cells (DC) but not by CD8+ DC. It is also expressed on monocytes, splenic plasmacytoid pre-DC, macrophages, and resident peritoneal cells. CD209a expression is downregulated upon cellular activation. CD209a is thought to play a role in the internalization and presentation of antigens to initiate immune responses.
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.
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