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Purified Mouse Anti-Human CD196 (CCR6)
Purified Mouse Anti-Human CD196 (CCR6)

Flow cytometric analysis of CD196 (CCR6) expression on human peripheral blood lymphocytes. Whole blood was stained with either Purified Mouse Anti-Human CD196 (CCR6) (Cat. No. 559560, black line histogram) or Purified Mouse IgG1, κ Isotype Control (Cat. No. 555746, grey line histogram). Secondary staining was performed with Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 553999), and visualization was carried out using PE Streptavidin (Cat. No. 554656). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing expression of CD196 (CCR6) (or Ig isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

Flow cytometric analysis of CD196 (CCR6) expression on human peripheral blood lymphocytes. Whole blood was stained with either Purified Mouse Anti-Human CD196 (CCR6) (Cat. No. 559560, black line histogram) or Purified Mouse IgG1, κ Isotype Control (Cat. No. 555746, grey line histogram). Secondary staining was performed with Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 553999), and visualization was carried out using PE Streptavidin (Cat. No. 554656). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing expression of CD196 (CCR6) (or Ig isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

製品詳細
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BD Pharmingen™
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)
0.5 mg/ml
IX 48
1235
AB_2291135
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


推奨アッセイ手順

The purified 11A9 antibody (Cat. No. 559560) can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes  (see image) and cell lines that express CCR6.

A multistep step staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of human CCR6 expression:

Step 1: Incubate the cells with 0.1 - 1 µg of purified 11A9 antibody at 4°C for 15-20 minutes. Wash cells two times with staining medium containing sodium azide (e.g., Dulbecco's PBS or tissue culture medium [without phenol red and biotin] with 0.09% sodium azide and 2% heat-inactivated FCS or 0.2% BSA).

Step 2: Incubate the cells with 0.25 µg of biotinylated goat anti-mouse Ig (Cat. No. 553999) at 4°C for 20 minutes. Wash cells two times.

Step 3: Incubate the cells with ≤ 0.06 mg of streptavidin-phycoerythrin (Cat. No. 554061) at 4°C for 20 minutes. Wash two times. Resuspend cells in staining medium and analyze stained cells with a BD FACScan™ Flow Cytometer (BD Biosciences, San Jose, CA) using appropriate specificity and compensation controls.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
559560 Rev. 4
抗体の詳細
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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.

559560 Rev. 4
フォーマットの詳細
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
559560 Rev.4
引用&参考文献
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Development References (7)

  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. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
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559560 Rev. 4

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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.