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    BB700 Mouse Anti-Human CD194
    BB700 Mouse Anti-Human CD194
    Multicolor flow cytometric analysis of CD194 expression on human peripheral blood lymphocytes. Human whole blood was treated with BD PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and then stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346/561005/561842; Top Plots) and BD Horizon™ BV421 Mouse Anti-Human CD8a (Cat. No. 562429/562428; Bottom Plots) antibodies, and either BD Horizon BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plots) or BD Horizon BB700 Mouse Anti-Human CD194 antibody (Cat. No. 566475/566476; Right Plots). Two-color flow cytometric dot plots showing the correlated expression patterns of CD194 (or Ig Isotype control staining) versus CD4 or CD8a were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometr9c analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
    BB700 Mouse Anti-Human CD194
    Multicolor flow cytometric analysis of CD194 expression on human peripheral blood lymphocytes. Human whole blood was treated with BD PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and then stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346/561005/561842; Top Plots) and BD Horizon™ BV421 Mouse Anti-Human CD8a (Cat. No. 562429/562428; Bottom Plots) antibodies, and either BD Horizon BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plots) or BD Horizon BB700 Mouse Anti-Human CD194 antibody (Cat. No. 566475/566476; Right Plots). Two-color flow cytometric dot plots showing the correlated expression patterns of CD194 (or Ig Isotype control staining) versus CD4 or CD8a were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometr9c analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
    Product Details
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    BD Horizon™
    CCR4; C-C chemokine receptor type 4; CMKBR4; K5-5
    Human (QC Testing)
    Mouse C57BL/6 IgG1, κ
    Human CCR4 Transfected Cell Line
    Flow cytometry (Routinely Tested)
    5 µl
    HCDM 2006
    1233
    AB_2744302
    Aqueous buffered solution containing ≤0.09% sodium azide.
    RUO


    Preparation And Storage

    Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.
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    Recommended Assay Procedures

         For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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 for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

         When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

         For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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    Product Notices

    1. 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).
    2. An isotype control should be used at the same concentration as the antibody of interest.
    3. 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.
    4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
    6. 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.
    7. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
    8. Cy is a trademark of GE Healthcare.
    9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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    566476 Rev. 1
    Antibody Details
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    1G1

    The 1G1 monoclonal antibody specifically binds to CD194, also known as the human CC Chemokine Receptor type 4 (CCR4).  CCR4 is expressed on activated Th2 cells, regulatory T cells, activated NK cells, basophils, monocytes and platelets.  CCR4 is a seven-transmembrane, G-protein-coupled receptor, and is the specific receptor for CC chemokines, CCL22/MDC/Macrophage-Derived Chemokine and CCL17/TARC/Thymus and Activation-Regulated Chemokine. It has been reported that CCR4 mRNA is expressed mainly in the thymus and spleen. The human CCR4 gene has been mapped to chromosome 3p24. The purified form of this antibody has been reported not to be a neutralizing antibody.  The immunogen used to generate the 1G1 hybridoma has been reported to be human CCR4 transfected L1.2 mouse lymphoma cells.

         The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

    566476 Rev. 1
    Format Details
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    BB700
    The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
    altImg
    BB700
    Blue 488 nm
    476 nm
    695 nm
    566476 Rev.1
    Citations & References
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    Development References (11)

    1. Andrew DP, Ruffing N, Kim CH, et al. C-C chemokine receptor 4 expression defines a major subset of circulating nonintestinal memory T cells of both Th1 and Th2 potential. J Immunol. 2000; 166(1):103-111. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition). View Reference
    2. Bonecchi R, Bianchi G, Bordignon PP, et al. Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s. J Exp Med. 1998; 187(1):129-134. (Biology). View Reference
    3. Campbell JJ, Haraldsen G, Pan J, et al. The chemokine receptor CCR4 in vascular recognition by cutaneous but not intestinal memory T cells. Nature. 1999; 400(6746):776-780. (Biology). View Reference
    4. D'Ambrosio D, Iellem A, Bonecchi R, et al. Selective up-regulation of chemokine receptors CCR4 and CCR8 upon activation of polarized human type 2 Th cells.. J Immunol. 1998; 161(10):5111-5. (Biology). View Reference
    5. Imai T, Baba M, Nishimura M, Kakizaki M, Takagi S, Yoshie O. The T cell-directed CC chemokine TARC is a highly specific biological ligand for CC chemokine receptor 4. J Biol Chem. 1997; 272(23):15036-15042. (Biology). View Reference
    6. Imai T, Chantry D, Raport CJ, et al. Macrophage-derived chemokine is a functional ligand for the CC chemokine receptor 4. J Biol Chem. 1998; 273(3):1764-1768. (Biology). View Reference
    7. Imai T, Nagira M, Takagi S, et al. Selective recruitment of CCR4-bearing Th2 cells toward antigen-presenting cells by the CC chemokines thymus and activation-regulated chemokine and macrophage-derived chemokine. Int Immunol. 1999; 11(1):81-88. (Biology). View Reference
    8. Power CA, Meyer A, Nemeth K, et al. Molecular cloning and functional expression of a novel CC chemokine receptor cDNA from a human basophilic cell line. J Biol Chem. 1995; 270(33):19495-19500. (Biology). View Reference
    9. Sallusto F, Lenig D, Mackay CR, Lanzavecchia A. Flexible programs of chemokine receptor expression on human polarized T helper 1 and 2 lymphocytes. J Exp Med. 1998; 187(6):875-883. (Biology). View Reference
    10. Samson M, Soularue P, Vassart G, Parmentier M. The genes encoding the human CC-chemokine receptors CC-CKR1 to CC-CKR5 (CMKBR1-CMKBR5) are clustered in the p21.3-p24 region of chromosome 3. Genomics. 1996; 36(3):522-526. (Biology). View Reference
    11. Zola H, Swart B, Banham A, et al. CD molecules 2006--human cell differentiation molecules.. J Immunol Methods. 2007; 319(1-2):1-5. (Clone-specific). View Reference
    View All (11) View Less
    566476 Rev. 1

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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.

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