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Two-color flow cytometric analysis of IL-17A expression in stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD4 antibody (Cat. No. 555349/561840/561841) and either BD Horizon™ BV786 Mouse IgG1 κ Isotype Control (Cat. No. 563330; Left Panel) or BD Horizon™ BV786 Mouse Anti-Human IL-17A antibody (Cat. No. 563745; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of IL-17A (or Ig Isotype control staining) versus CD4 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.


BD Horizon™ BV786 Mouse Anti-Human IL-17A

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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- BD Horizon Brilliant Violet 786 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- 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.
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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Human IL-17A, also known as IL-17, is a proinflammatory cytokine that is encoded by the IL17A gene in chromosome 6. IL-17A is produced as a disulfide-linked homodimer comprised of two mature 136-amino acid polypeptides. It is a member of the IL-17 family of structurally related cytokines, designated IL-17A through IL-17F. Activated memory T cells, especially Th17 cells (specialized IL-17A-producing CD4+ T cells distinct from Th1 and Th2 cells) produce IL-17 and provide protective immunity against pathogens. Activated CD8+ T cells, γδT cells, NK cells and neutrophils can also be activated to produce IL-17A. IL-17A binds to and exerts its biological activity through IL-17 receptors (IL-17R) that are expressed by a variety of target cells including fibroblasts, epithelial and endothelial cells, monocytes/macrophages and mast cells. The ubiquitous IL-17R expression pattern may explain the broad tissue responsiveness to IL-17. IL-17 induces stromal cells to secrete cytokines and chemokines involved in inflammatory and hematopoietic processes. For example, IL-17 induces fibroblasts to produce IL-6, IL-8, G-CSF and express increased surface ICAM-1. The N49-653 antibody reacts with human IL-17A.
The antibody was conjugated to BD Horizon BV786 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 786-nm. BD Horizon BV786 can be excited by the violet laser and detected in a filter used to detect Cy™7-like dyes (eg, 780/60-nm filter).

Development References (8)
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Boisvert M, Chetoui N, Gendron S, Aoudjit F. Alpha2beta1 integrin is the major collagen-binding integrin expressed on human Th17 cells. Eur J Immunol. 2010; 40(10):2710-2719. (Clone-specific: Flow cytometry). View Reference
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Fossiez F, Djossou O, Chomarat P, et al. T cell interleukin-17 induces stromal cells to produce proinflammatory and hematopoietic cytokines. J Exp Med. 1996; 183(6):2593-2603. (Biology). View Reference
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Korn T, Oukka M, Kuchroo V, Bettelli E. Th17 cells: effector T cells with inflammatory properties. Semin Immunol. 2007; 19(6):362-371. (Biology). View Reference
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Melton AC, Melrose J, Alajoki L, et al. Regulation of IL-17A production is distinct from IL-17F in a primary human cell co-culture model of T cell-mediated B cell activation. PLoS ONE. 2013; 8(3):e58966. (Clone-specific: Flow cytometry). View Reference
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Moseley TA, Haudenschild DR, Rose L, Reddi AH. Interleukin-17 family and IL-17 receptors. Cytokine Growth Factor Rev. 2003; 14(2):155-174. (Biology). View Reference
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Weaver CT, Hatton RD, Mangan PR, Harrington LE. IL-17 family cytokines and the expanding diversity of effector T cell lineages. Annu Rev Immunol. 2007; 25:821-852. (Biology). View Reference
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Yao Z, Painter SL, Fanslow WC, et al. Human IL-17: a novel cytokine derived from T cells. J Immunol. 1995; 155(12):5483-5486. (Biology). View Reference
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Yao Z, Spriggs MK, Derry JM, et al. Molecular characterization of the human interleukin (IL)-17 receptor. Cytokine. 1997; 9(11):794-800. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.