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    BV650 Mouse Anti-Human IL-17F
    BV650 Mouse Anti-Human IL-17F
    Multicolor flow cytometric analysis of IL-17F expression by resting and activated human peripheral blood CD4+ T cells. Human peripheral blood mononuclear cells were either unstimulated (Upper Panels) or stimulated (Lower Panels) with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139, 50 ng/ml) plus Ionomycin (Sigma I-0634, 1 μg/ml) in the presence of  BD GolgiStop™  Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) for 5 hours. The cells were 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 PerCP-Cy™5.5 Mouse Anti-Human CD4 (Cat. No. 560650), BD Horizon BV650 Mouse Anti-Human IL-17F (Cat. No. 564264), PE Mouse Anti-Human IL-17A (Cat. No. 560486/560487; Left Panels), and FITC Mouse Anti-Human IFN-γ (Cat. No. 554700/552887/561053; Right Panels) antibodies. Two-color flow cytometric contour plots showing the correlated expression of IL-17F versus IL-17A or IFN-γ were derived from CD4-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.
    BV650 Mouse Anti-Human IL-17F
    Multicolor flow cytometric analysis of IL-17F expression by resting and activated human peripheral blood CD4+ T cells. Human peripheral blood mononuclear cells were either unstimulated (Upper Panels) or stimulated (Lower Panels) with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139, 50 ng/ml) plus Ionomycin (Sigma I-0634, 1 μg/ml) in the presence of  BD GolgiStop™  Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) for 5 hours. The cells were 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 PerCP-Cy™5.5 Mouse Anti-Human CD4 (Cat. No. 560650), BD Horizon BV650 Mouse Anti-Human IL-17F (Cat. No. 564264), PE Mouse Anti-Human IL-17A (Cat. No. 560486/560487; Left Panels), and FITC Mouse Anti-Human IFN-γ (Cat. No. 554700/552887/561053; Right Panels) antibodies. Two-color flow cytometric contour plots showing the correlated expression of IL-17F versus IL-17A or IFN-γ were derived from CD4-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.
    Product Details
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    BD Horizon™
    Interleukin-17F; ML-1; cytokine ML-1
    Human (QC Testing)
    Mouse IgG1, κ
    Human IL-17F
    Intracellular staining (flow cytometry) (Routinely Tested)
    5 µl
    AB_2869555
    Aqueous buffered solution containing BSA and ≤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™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 were removed.
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    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).

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    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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
    5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
    6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    7. BD Horizon Brilliant Violet 650 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.
    8. 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.
    9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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    564264 Rev. 2
    Antibody Details
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    O33-782

    The O33-782 monoclonal antibody specifically binds to Interleukin-17F (IL-17F). IL-17F is a member of the IL-17 family of cytokines. IL-17F is encoded by the IL17F gene located in chromosome 6 (location: 6p12). IL-17F is a proinflammatory cytokine that is produced by activated T cells including differentiated CD4+ T helper 17 (Th17) cells. Activated Th17 cells can express disulfide-linked IL-17F and IL-17A homodimers as well as IL-17A/IL-17F heterodimers. These IL-17 dimers act by binding to and signaling through IL-17 receptor complexes (IL-17R). IL-17R are comprised of transmembrane IL-17RA and IL-17-RC protein subunits that are expressed by a variety of target cells including epithelial and endothelial cells, keratinocytes, fibroblasts, and granulocytes. IL-17F can induce target cells to produce proinflammatory cytokines such as IL-1β, IL-6, G-CSF, GM-CSF, and TNF and chemokines including CXCL1/Gro-α, CXCL2/Gro-β, and CXCL8/IL-8 that attract and activate leukocytes, eg, neutrophils. Th17 and other IL-17F-producing cells play protective roles in the clearance of extracellular pathogens, including bacteria and fungi. IL-17F can also play adverse roles in inflammation associated with asthma and autoimmune diseases.

    The antibody was conjugated to BD Horizon BV650 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 650-nm.  BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter).  Due to the excitation and emission characteristics of the acceptor dye, there will be  spillover into the APC and Alexa Fluor® 700 detectors.  However, the spillover can be corrected through compensation as with any other dye combination.

    564264 Rev. 2
    Format Details
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    BV650
    The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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    BV650
    Violet 405 nm
    406 nm
    649 nm
    564264 Rev.2
    Citations & References
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    Development References (7)

    1. Fouser LA, Wright JF, Dunussi-Joannopoulos K, Collins M. Th17 cytokines and their emerging roles in inflammation and autoimmunity. Immunol Rev. 2008; 226:87-102. (Biology). View Reference
    2. 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
    3. Shen F, Gaffen SL. Structure-function relationships in the IL-17 receptor: implications for signal transduction and therapy. Cytokine. 2008; 41(2):92-104. (Biology). View Reference
    4. Starnes T, Robertson MJ, Sledge G, et al.. Cutting edge: IL-17F, a novel cytokine selectively expressed in activated T cells and monocytes, regulates angiogenesis and endothelial cell cytokine production. J Immunol. 2001; 167(8):4137-4140. (Biology). View Reference
    5. Wang YH, Liu YJ. The IL-17 cytokine family and their role in allergic inflammation. Curr Opin Immunol. 2008; 20(6):697-702. (Biology). View Reference
    6. Wright JF, Bennett F, Li B, et al. The human IL-17F/IL-17A heterodimeric cytokine signals through the IL-17RA/IL-17RC receptor complex. J Immunol. 2008; 181(4):2799-2805. (Biology). View Reference
    7. Wright JF, Guo Y, Quazi A, et al. Identification of an interleukin 17F/17A heterodimer in activated human CD4+ T cells. J Biol Chem. 2007; 282(18):13447-13455. (Biology). View Reference
    View All (7) View Less
    564264 Rev. 2

    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.

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