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BB700 Mouse Anti-Human CD196 (CCR6)
BB700 Mouse Anti-Human CD196 (CCR6)
Multiparameter flow cytometric analysis of CD196 (CCR6) expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD196 (CCR6) antibody (Cat. No. 566477/566478; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric dot plots showing the correlated expression of CD196 (CCR6) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Multiparameter flow cytometric analysis of CD196 (CCR6) expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD196 (CCR6) antibody (Cat. No. 566477/566478; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric dot plots showing the correlated expression of CD196 (CCR6) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. 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_2744303
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.

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.

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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566477 Rev. 1
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 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.

566477 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.
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BB700
Blue 488 nm
476 nm
695 nm
566477 Rev.1
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. Cabezon R, Sintes J, Llinas L, Benitez-Ribas D. Analysis of HLDA9 mAbs on plasmacytoid dendritic cell. Immunol Lett. 2011; 134(2):167-173. (Clone-specific: Flow cytometry). View Reference
  4. 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
  5. 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
  6. 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
  7. 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
  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. Llinas L, Lazaro A, de Salort J, Matesanz-Isabel J, Sintes J, Engel P. Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry. Immunol Lett. 2011; 134(2):113-121. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
  12. Ramos-Medina R, Montes-Moreno S, Maestre L, et al. Immunohistochemical analysis of HLDA9 Workshop antibodies against cell-surface molecules in reactive and neoplastic lymphoid tissues. Immunol Lett. 2011; 134(2):150-156. (Clone-specific: Immunohistochemistry). View Reference
  13. 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
  14. 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
  15. 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
View All (15) View Less
566477 Rev. 1

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