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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 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™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.
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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.