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      BB515 Mouse Anti-Human CD196 (CCR6)
      BB515 Mouse Anti-Human CD196 (CCR6)
      Flow cytometric analysis of CD196 (CCR6) expression on human peripheral blood cells. Human whole blood was stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD196 (CCR6) antibody (Cat. No. 564479; solid 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 LSRFortessa™ Cell Analyzer System.
      BB515 Mouse Anti-Human CD196 (CCR6)
      Flow cytometric analysis of CD196 (CCR6) expression on human peripheral blood cells. Human whole blood was stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD196 (CCR6) antibody (Cat. No. 564479; solid 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 LSRFortessa™ Cell Analyzer System.
      Product Details
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      BD Horizon™
      BN-1; C-C CKR-6; C-C chemokine receptor type 6; CC-CKR-6; CCR-6
      Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
      Mouse IgG1, κ
      Human CD196/CCR6 Peptide
      Flow cytometry (Routinely Tested)
      5 µl
      IX 48
      1235
      AB_2738825
      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™ BB515 under optimum conditions and unconjugated antibody was removed.
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      Recommended Assay Procedures

        BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

      For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

      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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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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. 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.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564479 Rev. 4
      Antibody Details
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      11A9

       The 11A9 monoclonal antibody specifically binds to CD196, which is also known as CCR6. CCR6 is a seven-transmembrane, G-protein-coupled, glycoprotein receptor that is a member of the beta chemokine receptor family. The human CCR6 gene has been mapped to chromosome 6q27. CCR6 is a receptor for the CC chemokine CCL20/MIP-3alpha/LARC/Exodus and also binds with lower affinity to and mediates responses to beta-defensin2/hBD-2. CCR6 is predominantly expressed by B lymphocytes, certain subsets of effector and memory T cells and by immature dendritic cells but not by monocytes, NK cells, or granulocytes. Skin-homing CLA (Cutaneous Lymphocyte Antigen)-positive memory T cells, Th1 cells, regulatory T cells and IL-17A-producing Th17 cells predominantly express high levels of CCR6. CCR6 mediates the trafficking of T, B, and dendritic cells to epithelial sites near the skin and mucosal surfaces during inflammatory and immunological responses. An N-terminal peptide of human CCR6 was used as an immunogen to generate the 11A9 hybridoma. The 11A9 antibody does not cross-react with human CCR1, CCR2, CCR3, CCR4, CCR5, CCR7, CCR8, CCR9, CXCR1, CXCR2, CXCR3, CXCR4 and CXCR5 receptors. This antibody is NOT a neutralizing antibody.

      The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

      564479 Rev. 4
      Format Details
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      BB515
      The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BB515
      Blue 488 nm
      490 nm
      515 nm
      564479 Rev.4
      Citations & References
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      Development References (15)

      1. Baba M, Imai T, Nishimura M, et al. Identification of CCR6, the specific receptor for a novel lymphocyte-directed CC chemokine LARC. J Biol Chem. 1997; 272(23):14893-14898. (Biology). View Reference
      2. Brandes M, Willimann K, Lang AB, et al. Flexible migration program regulates gamma delta T-cell involvement in humoral immunity. Blood. 2003; 102(10):3693-3701. (Clone-specific: Flow cytometry). View Reference
      3. Greaves DR, Wang W, Dairaghi DJ, et al. CCR6, a CC chemokine receptor that interacts with macrophage inflammatory protein 3alpha and is highly expressed in human dendritic cells. J Exp Med. 1997; 186(6):837-844. (Biology). View Reference
      4. Homey B, Dieu-Nosjean MC, Wiesenborn A, et al. Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis. J Immunol. 2000; 164(12):6621-6632. (Biology). View Reference
      5. Kim CH, Rott L, Kunkel EJ, et al. Rules of chemokine receptor association with T cell polarization in vivo. J Clin Invest. 2001; 108(9):1331-1339. (Biology). View Reference
      6. Liao F, Alderson R, Su J, Ullrich SJ, Kreider BL, Farber JM. STRL22 is a receptor for the CC chemokine MIP-3alpha. Biochem Biophys Res Commun. 1997; 236(1):212-217. (Biology). View Reference
      7. Liao F, Lee HH, Farber JM. Cloning of STRL22, a new human gene encoding a G-protein-coupled receptor related to chemokine receptors and located on chromosome 6q27. Genomics. 1997; 40(1):175-180. (Biology). View Reference
      8. Liao F, Rabin RL, Smith CS, Sharma G, Nutman TB, Farber JM. CC-chemokine receptor 6 is expressed on diverse memory subsets of T cells and determines responsiveness to macrophage inflammatory protein 3 alpha. J Immunol. 1999; 162(1):186-194. (Biology). View Reference
      9. Liao F, Shirakawa AK, Foley JF, Rabin RL, Farber JM. Human B cells become highly responsive to macrophage-inflammatory protein-3 alpha/CC chemokine ligand-20 after cellular activation without changes in CCR6 expression or ligand binding. J Immunol. 2002; 168(10):4871-4880. (Clone-specific: Flow cytometry). View Reference
      10. Lim HW, Lee J, Hillsamer P, Kim CH. Human Th17 cells share major trafficking receptors with both polarized effector T cells and FOXP3+ regulatory T cells. J Immunol. 2008; 180(1):122-129. (Biology). View Reference
      11. Power CA, Church DJ, Meyer A, et al. Cloning and characterization of a specific receptor for the novel CC chemokine MIP-3alpha from lung dendritic cells. J Exp Med. 1997; 186(6):825-835. (Biology). View Reference
      12. Sallusto F, Lenig D, Forster R, Lipp M, Lanzavecchia A. Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature. 1999; 401(6754):708-712. (Clone-specific: Flow cytometry). View Reference
      13. Thomas SY, Banerji A, Medoff BD, Lilly CM, Luster AD. Multiple chemokine receptors, including CCR6 and CXCR3, regulate antigen-induced T cell homing to the human asthmatic airway. J Immunol. 2007; 179(3):1901-1912. (Clone-specific: Flow cytometry). View Reference
      14. Yang D, Chertov O, Bykovskaia SN, et al. Beta-defensins: linking innate and adaptive immunity through dendritic and T cell CCR6. Science. 1999; 286(5439):525-528. (Biology). View Reference
      15. Zaballos A, Varona R, Gutierrez J, Lind P, Marquez G. Molecular cloning and RNA expression of two new human chemokine receptor-like genes. Biochem Biophys Res Commun. 1996; 227(3):846-853. (Biology). View Reference
      View All (15) View Less
      564479 Rev. 4

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      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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