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Flow cytometric analysis of CD183 (CXCR3) expression on human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ BV711 Mouse anti-Human CD183 antibody (Cat. No. 563156; solid line histogram) or with a BD Horizon™BV711 Mouse IgG1, κ Isotype Control (Cat. No. 563044; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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.
Flow cytometric analysis of CD183 (CXCR3) expression on human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ BV711 Mouse anti-Human CD183 antibody (Cat. No. 563156; solid line histogram) or with a BD Horizon™BV711 Mouse IgG1, κ Isotype Control (Cat. No. 563044; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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.
Flow cytometric analysis of CD183 (CXCR3) expression on human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ BV711 Mouse anti-Human CD183 antibody (Cat. No. 563156; solid line histogram) or with a BD Horizon™BV711 Mouse IgG1, κ Isotype Control (Cat. No. 563044; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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.
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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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
The 1C6/CXCR3 monoclonal antibody specifically binds to human CD183, also known as the CXCR3 chemokine receptor. CD183 is a 40-41 kDa seven-transmembrane protein and member of the G protein-coupled receptor family. CD183 is expressed primarily on activated T cells that infiltrate inflammatory sites. It has also been detected on some circulating T cells, B cells, and NK cells. Reports show that some CXCR3-positive T cells also express CCR5 and are mostly CD45RO-positive cells. Three ligands for CXCR3 have been identified. They are CXCL9 (Mig/monokine induced by interferon-γ), CXCL10 (IP-10/interferon-γ inducible 10-kD protein), and CXCL11 (I-TAC/interferon-inducible T-cell alpha chemoattractant). These chemokines are produced by a variety of cells upon stimulation by IFN-γ and interact with CXCR3 to mediate T-cell chemotaxis. This reagent has been reported to be suitable for immunohistochemical staining of acetone-fixed, frozen sections and/or formalin-fixed, paraffin-embedded tissue sections with citrate pretreatment. Clone 1C6/CXCR3 also cross reacts with a subset of peripheral blood lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys. The distribution of lymphocytes is similar to that observed with CD183-positive peripheral blood lymphocytes from normal human donors. CXCR3 has been clustered as CD183 in the VIIth HLDA workshop.
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-Cy™5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
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