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      PerCP-Cy™5.5 Mouse anti-Human IL-17F
      PerCP-Cy™5.5 Mouse anti-Human IL-17F
      Flow cytometric analysis of IL-17F expression by resting or activated human peripheral blood mononuclear cells (PBMC). Human PBMC were either unstimulated (Left Panels) or stimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 5 hours (Middle Panels) or cultured in Th17 polarization conditions and restimulated with PMA and Ionomycin in the presence of BD GolgiStop™ for 5 hours (Right Panels). Cells were then fixed and permeabilized using BD Cytofix/Cytoperm™ reagents (Cat. No. 554714) followed by staining with PerCP-Cy™5.5 Mouse anti-Human IL-17F (Cat. No. 561335), PE Mouse anti-Human CD4 (Cat. No. 555347), FITC Mouse Anti-Human IFN-γ (Cat. No. 554700) and Alexa Fluor® 647 Mouse anti-Human IL-17A (Cat. No. 560490). Two-color flow cytometric dot plots showing the correlated expression patterns 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 cytometry was performed using a BD™ LSR II Flow Cytometry System. Other compatible fixation and permeabilization treatments are listed in the \"Recommended Assay Procedure.\"
      PerCP-Cy™5.5 Mouse anti-Human IL-17F
      Flow cytometric analysis of IL-17F expression by resting or activated human peripheral blood mononuclear cells (PBMC). Human PBMC were either unstimulated (Left Panels) or stimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 5 hours (Middle Panels) or cultured in Th17 polarization conditions and restimulated with PMA and Ionomycin in the presence of BD GolgiStop™ for 5 hours (Right Panels). Cells were then fixed and permeabilized using BD Cytofix/Cytoperm™ reagents (Cat. No. 554714) followed by staining with PerCP-Cy™5.5 Mouse anti-Human IL-17F (Cat. No. 561335), PE Mouse anti-Human CD4 (Cat. No. 555347), FITC Mouse Anti-Human IFN-γ (Cat. No. 554700) and Alexa Fluor® 647 Mouse anti-Human IL-17A (Cat. No. 560490). Two-color flow cytometric dot plots showing the correlated expression patterns 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 cytometry was performed using a BD™ LSR II Flow Cytometry System. Other compatible fixation and permeabilization treatments are listed in the \"Recommended Assay Procedure.\"
      Product Details
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      BD Pharmingen™
      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_10613817
      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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
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      Recommended Assay Procedures

      This antibody conjugate is suitable for intracellular staining of human peripheral blood mononuclear cells using BD Cytofix/Cytoperm™ reagents, BD Pharmingen™ Human FoxP3 Buffer Set or BD™ Phosflow fixation and permeabilization buffers (Fix buffer I with Perm/Wash Buffer I, Perm Buffer II, or Perm Buffer III).

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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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. 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.
      5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
      6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
      7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Cy is a trademark of GE Healthcare.
      10. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      561336 Rev. 3
      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.

      561336 Rev. 3
      Format Details
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      PerCP-Cy5.5
      PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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.
      altImg
      PerCP-Cy5.5
      Blue 488 nm
      482 nm
      676 nm
      561336 Rev.3
      Citations & References
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      Development References (6)

      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. 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
      3. 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
      4. 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
      5. 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
      6. 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 (6) View Less
      561336 Rev. 3

      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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