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Flow cytometric analysis of CD183 (CXCR3) expression on human peripheral blood lymphocytes. Whole blood was stained with either BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat. No. 562652; dashed line histogram) or BD Horizon™ BV605 Mouse Anti-Human CD183 antibody (Cat. No. 564032; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV605 Mouse Anti-Human CD183
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
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).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- CF™ is a trademark of Biotium, Inc.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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.
This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
Development References (5)
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Loetscher M, Gerber B, Loetscher P, et al. Chemokine receptor specific for IP10 and mig: structure, function, and expression in activated T-lymphocytes. J Exp Med. 1996; 184(3):963-969. (Biology). View Reference
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Marcher C, Moller BK, Lillevang ST, Kristensen T. CXCR4 and IL17R are downregulated on cord-blood CD34-positive cells during short-term culture. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:629-632.
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Piali L, Weber C, LaRosa G, et al. The chemokine receptor CXCR3 mediates rapid and shear-resistant adhesion-induction of effector T lymphocytes by the chemokines IP10 and Mig. Eur J Immunol. 1998; 28(3):961-972. (Biology). View Reference
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Qin S, Rottman JB, Myers P, et al. The chemokine receptors CXCR3 and CCR5 mark subsets of T cells associated with certain inflammatory reactions. J Clin Invest. 1998; 101(4):746-754. (Immunogen: Blocking, Flow cytometry, Immunohistochemistry, Induction). View Reference
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Uguccioni M, Willimann K. Cytokine/Chemokine Receptors: Section report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:237-243.
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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.