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    BV650 Rat Anti-Mouse IL-17A
    BV650 Rat Anti-Mouse IL-17A
    Flow cytometric analysis of IL-17A-producing mouse EL-4 cells. Cells from the mouse EL4 (T lymphoma, ATCC TIB-39) cell line were stimulated (5 hr) with Phorbol 12-Myristate 13-Acetate (PMA; 50 ng/ml final concentration; Sigma, Cat. No. P-8139) and Ionomycin (1000 ng/ml final concentration; Sigma, Cat. No. I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), 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 BD Horizon™ BV650 Rat IgG1 κ Isotype Control (Cat. No. 563848; Left Panel) or BD Horizon BV650 Rat Anti-Mouse IL-17A antibody (Cat. No. 564170; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing correlated expression patterns of IL-17A (or Ig Isotype control staining) versus cellular autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
    BV650 Rat Anti-Mouse IL-17A
    Flow cytometric analysis of IL-17A-producing mouse EL-4 cells. Cells from the mouse EL4 (T lymphoma, ATCC TIB-39) cell line were stimulated (5 hr) with Phorbol 12-Myristate 13-Acetate (PMA; 50 ng/ml final concentration; Sigma, Cat. No. P-8139) and Ionomycin (1000 ng/ml final concentration; Sigma, Cat. No. I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), 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 BD Horizon™ BV650 Rat IgG1 κ Isotype Control (Cat. No. 563848; Left Panel) or BD Horizon BV650 Rat Anti-Mouse IL-17A antibody (Cat. No. 564170; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing correlated expression patterns of IL-17A (or Ig Isotype control staining) versus cellular autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
    Product Details
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    BD Horizon™
    IL-17; Il17; IL-17A; Il17a; CTLA-8; Ctla8; Interleukin-17A; interleukin 17A
    Mouse (QC Testing)
    Rat LEW, also known as Lewis IgG1, κ
    Recombinant Mouse IL-17A Protein
    Intracellular staining (flow cytometry) (Routinely Tested)
    0.2 mg/ml
    AB_2738641
    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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    Product Notices

    1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
    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. Brilliant Violet™ 650 is a trademark of Sirigen.
    7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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    564170 Rev. 1
    Antibody Details
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    TC11-18H10

    The TC11-18H10 monoclonal antibody specifically binds to recombinant and natural mouse IL-17A proteins. IL-17A, also known as CTLA-8, is a T cell-derived cytokine that promotes inflammatory responses.  Mouse IL-17A is a proinflammatory cytokine that can induce the release of IL-6 by mouse stromal cells. It has been shown to support the growth of hemopoietic progenitors in vitro; it can also stimulate granulopoiesis in vivo. The TC11-18H10 antibody has been reported to neutralize IL-17A activity.  Recent studies have shown that IL-17A is produced by a unique subset of Th17 cells that develop along a pathway distinct from the Th1- and Th2- cell differentiation pathways. The mouse IL-17A cDNA was isolated from a cDNA library generated from TCRαβ+CD4-CD8- thymocytes.

    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.

    564170 Rev. 1
    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
    564170 Rev.1
    Citations & References
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    Development References (4)

    1. Coquet JM, Chakravarti S, Kyparissoudis K, et al. Diverse cytokine production by NKT cell subsets and identification of an IL-17-producing CD4-NK1.1- NKT cell population. Proc Natl Acad Sci U S A. 2008; 105(32):11287-11292. (Clone-specific: Flow cytometry). View Reference
    2. Dong C. Th17 cells: Current understanding of their generation and regulation. Eur J Immunol. 2009; 39(3):640-644. (Biology). View Reference
    3. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Flow cytometry). View Reference
    4. Schwarzenberger P, La Russa V, Miller A, et al. IL-17 stimulates granulopoiesis in mice: use of an alternate, novel gene therapy-derived method for in vivo evaluation of cytokines. J Immunol. 1998; 161(11):6383-6389. (Biology). View Reference
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    564170 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.

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