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BD Pharmingen™ PE Mouse Anti-Human CD196 (CCR6)
Clone 11A9 (RUO)
Flow cytometric analysis of CCR6 expression on stimulated human PBMC. PBMC were stimulated with PMA/Ionomycin in the presence of BD GolgiStop™ (Cat. No. 554724) for 5 hours. After stimulation, cells were surface stained with PE Mouse Anti-Human CD196 (CCR6) (Cat. No. 559562/561019/551773), fixed and permeabilized using BD Cytofix/Cytoperm™ reagents (Cat. No. 554714), and intracellular stained with either FITC anti-human IFN-γ (Cat. No. 554700; left panel), APC anti-human IL-4 (Cat. No.554486; center panel), or Alexa Fluor® 647 anti-human IL-17A (Cat. No. 560437; right panel). The dot plots were derived gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of CCR6 expression on stimulated human PBMC. PBMC were stimulated with PMA/Ionomycin in the presence of BD GolgiStop™ (Cat. No. 554724) for 5 hours. After stimulation, cells were surface stained with PE Mouse Anti-Human CD196 (CCR6) (Cat. No. 559562/561019/551773), fixed and permeabilized using BD Cytofix/Cytoperm™ reagents (Cat. No. 554714), and intracellular stained with either FITC anti-human IFN-γ (Cat. No. 554700; left panel), APC anti-human IL-4 (Cat. No.554486; center panel), or Alexa Fluor® 647 anti-human IL-17A (Cat. No. 560437; right panel). The dot plots were derived gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of CCR6 expression on stimulated human PBMC. PBMC were stimulated with PMA/Ionomycin in the presence of BD GolgiStop™ (Cat. No. 554724) for 5 hours. After stimulation, cells were surface stained with PE Mouse Anti-Human CD196 (CCR6) (Cat. No. 559562/561019/551773), fixed and permeabilized using BD Cytofix/Cytoperm™ reagents (Cat. No. 554714), and intracellular stained with either FITC anti-human IFN-γ (Cat. No. 554700; left panel), APC anti-human IL-4 (Cat. No.554486; center panel), or Alexa Fluor® 647 anti-human IL-17A (Cat. No. 560437; right panel). The dot plots were derived gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of CCR6 expression on stimulated human PBMC. PBMC were stimulated with PMA/Ionomycin in the presence of BD GolgiStop™ (Cat. No. 554724) for 5 hours. After stimulation, cells were surface stained with PE Mouse Anti-Human CD196 (CCR6) (Cat. No. 559562/561019/551773), fixed and permeabilized using BD Cytofix/Cytoperm™ reagents (Cat. No. 554714), and intracellular stained with either FITC anti-human IFN-γ (Cat. No. 554700; left panel), APC anti-human IL-4 (Cat. No.554486; center panel), or Alexa Fluor® 647 anti-human IL-17A (Cat. No. 560437; right panel). The dot plots were derived gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ System.
Product Details
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)
0.2 mg/ml
IX 48
AB_397273
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Recommended Assay Procedures
The PE-conjugated 11A9 antibody (Cat. No. 559562/561019/551773) can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes (see image) and cell lines that express CCR6. Please titrate between 0.1 and 1.0 µg/test.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
Fixation/Permeabilization Kit RUO
Size
250 Tests
Cat No.
554714
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 ML
Cat No.
554724
559562 Rev. 5
Antibody Details
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.
559562 Rev. 5
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
496 nm, 566 nm
576 nm
559562 Rev.5
Citations & References
View product citations for antibody "559562" on CiteAb
Development References (7)
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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
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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
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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
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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
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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
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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
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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
559562 Rev. 5
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.