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      R718 Mouse Anti-Human IL-17A
      R718 Mouse Anti-Human IL-17A
      Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 μg/ml for plate coating) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 2 μg/ml) antibodies and Recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml) and Human IL-4 (Cat. No. 554605; 25 ng/ml) proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate (PMA; Sigma, Cat. No. P-8139; 50 ng/ml) and Ionomycin (Sigma, Cat. No. I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724).        The cells were harvested and washed with Stain Buffer (FBS) (Cat. No. 554656) and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat No. 564724), FITC Mouse Anti-Human CD45RO  (Cat. No. 555492; Left Plot), BD Horizon™ BV421 Mouse Anti-Human IL-4 (Cat. No.  564110; Middle Plot Plot), and PE Mouse Anti-Human IFN-γ (Cat. No. 559326; Right Plot Plot) antibodies and with either BD Horizon™ R718 Rat IgG1 κ Isotype Control (Cat. No. 566928; Data Not Shown) or BD Horizon™ R718 Mouse Anti-Human IL-17 antibody (Cat. No. 567094/567247) using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-γ were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
      R718 Mouse Anti-Human IL-17A
      Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 μg/ml for plate coating) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 2 μg/ml) antibodies and Recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml) and Human IL-4 (Cat. No. 554605; 25 ng/ml) proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate (PMA; Sigma, Cat. No. P-8139; 50 ng/ml) and Ionomycin (Sigma, Cat. No. I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724).        The cells were harvested and washed with Stain Buffer (FBS) (Cat. No. 554656) and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat No. 564724), FITC Mouse Anti-Human CD45RO  (Cat. No. 555492; Left Plot), BD Horizon™ BV421 Mouse Anti-Human IL-4 (Cat. No.  564110; Middle Plot Plot), and PE Mouse Anti-Human IFN-γ (Cat. No. 559326; Right Plot Plot) antibodies and with either BD Horizon™ R718 Rat IgG1 κ Isotype Control (Cat. No. 566928; Data Not Shown) or BD Horizon™ R718 Mouse Anti-Human IL-17 antibody (Cat. No. 567094/567247) using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-γ were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
      Product Details
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      BD Horizon™
      IL-17; IL-17A; CTLA8; Cytotoxic T-lymphocyte-associated serine esterase 8
      Human (QC Testing)
      Mouse IgG1, κ
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      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 to the dye under optimum conditions and unreacted dye 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 BD 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 BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. 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).
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
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      567681 Rev. 1
      Antibody Details
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      SCPL1362

      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 SCPL1362 antibody reacts with human IL-17A.

      567681 Rev. 1
      Format Details
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      R718
      The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      R718
      Red 627-640 nm
      695 nm
      718 nm
      567681 Rev.1
      Citations & References
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      View product citations for antibody "567681" on CiteAb

      Development References (6)

      1. 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
      2. 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
      3. 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
      4. 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
      5. 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
      6. 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
      View All (6) View Less
      567681 Rev. 1

      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.