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    BV421 Mouse Anti-Human IL-9
    BV421 Mouse Anti-Human IL-9
    Flow cytometric analysis of IL-9 expressed in stimulated human lymphocytes. PBMC were cultured (5 d)  with plate-bound NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 μg/ml, coated overnight at 4°C) and soluble NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 1 μg/ml) antibodies plus recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml), IL-4 (Cat. No. 554605; 50 ng/ml), and TGF-β (Corning; Cat. No. 354039; 10 ng/ml) proteins and NA/LE Mouse Anti-Human IFN-γ antibody (Cat. No. 554698; 10 μg/ml). The cells were restimulated (5 h) with PMA (Sigma P8139; 50 ng/ml) and ionomycin (Sigma I9657; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The cells were washed, and then fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeablization Kit (Cat. No. 554714) and stained with BD Horizon™ BV421 Mouse Anti-Human IL-9 antibody (Cat. No. 564254). A two-parameter flow cytometric contour plot showing the coexpressed levels of IL-9 versus Autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRII™ System.
    BV421 Mouse Anti-Human IL-9
    Flow cytometric analysis of IL-9 expressed in stimulated human lymphocytes. PBMC were cultured (5 d)  with plate-bound NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 μg/ml, coated overnight at 4°C) and soluble NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 1 μg/ml) antibodies plus recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml), IL-4 (Cat. No. 554605; 50 ng/ml), and TGF-β (Corning; Cat. No. 354039; 10 ng/ml) proteins and NA/LE Mouse Anti-Human IFN-γ antibody (Cat. No. 554698; 10 μg/ml). The cells were restimulated (5 h) with PMA (Sigma P8139; 50 ng/ml) and ionomycin (Sigma I9657; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The cells were washed, and then fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeablization Kit (Cat. No. 554714) and stained with BD Horizon™ BV421 Mouse Anti-Human IL-9 antibody (Cat. No. 564254). A two-parameter flow cytometric contour plot showing the coexpressed levels of IL-9 versus Autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRII™ System.
    Product Details
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    BD Horizon™
    IL9; IL-9; interleukin-9; HP40; P40
    Human (QC Testing)
    Mouse C57BL/6 IgG1, κ
    Human IL-9 Recombinant Protein
    Intracellular staining (flow cytometry) (Routinely Tested)
    5 µl
    AB_2738706
    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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
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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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
    6. Brilliant Violet™ 421 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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    564254 Rev. 1
    Antibody Details
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    MH9A3

    The MH9A3 monoclonal antibody specifically binds to human interleukin-9 (IL-9). Human IL-9 is a multifunctional cytokine and a member of the type I cytokine (hematopoietin) family that includes IL-2, IL-4, IL-7, IL-15 and IL-21. This cytokine is encoded by the IL9 gene that is resident on chromosome 5q31.1. IL-9 is expressed by activated CD4-positive T helper cells, by some transformed T cells and by eosinophils, mast cells and neutrophils. IL-9 induces the proliferation, differentiation, and effector function of various cell types including T lymphocytes, B lymphocytes, mast cells, eosinophils, neutrophils, hematopoietic cells and epithelial cells. It potentiates the interleukin-4-induced IgM, IgG and IgE responses by human B lymphocytes. IL-9 has been implicated in human allergic disorders such as asthma and malignancies such as Hodgkin's disease. IL-9 exerts its biological activities through binding to the surface IL-9 receptor (IL-9R) complex comprised of the IL-9R alpha subunit (IL-9Rα; CD129) and the common cytokine receptor gamma subunit (γc; CD132). IL-9 signaling through its receptor includes activation of the Janus kinases 1 and 3 ( JAK1 and JAK3) and activation of Signal transducer and activator of transcription 1, 3 and 5 factors (STAT1, STAT3 and STAT5).

    The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

    564254 Rev. 1
    Format Details
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    BV421
    The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
    altImg
    BV421
    Violet 405 nm
    407 nm
    423 nm
    564254 Rev.1
    Citations & References
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    Development References (9)

    1. Demoulin JB, Van Roost E, Stevens M, Groner B, Renauld JC. Distinct roles for STAT1, STAT3, and STAT5 in differentiation gene induction and apoptosis inhibition by interleukin-9. J Biol Chem. 1999; 274:25855-25861. (Biology). View Reference
    2. Dugas B, Renauld JC, Pène J, et al. Interleukin-9 potentiates the interleukin-4-induced immunoglobulin (IgG, IgM and IgE) production by normal human B lymphocytes. Eur J Immunol. 1993 July; 23(7):1687-1692. (Biology). View Reference
    3. Houssiau FA, Schandene L, Stevens M. A cascade of cytokines is responsible for IL-9 expression in human T cells. Involvement of IL-2, IL-4, and IL-10. J Immunol. 1995; 154(6):2624-2630. (Biology). View Reference
    4. Jenmalm MC, Van Snick J, Cormont F, Salman B. Allergen-induced Th1 and Th2 cytokine secretion in relation to specific allergen sensitization and atopic symptoms in children. Clin Exp Allergy. 2001 October; 31(10):1528-1535. (Immunogen: ELISA). View Reference
    5. Knoops L, Renauld JC. IL-9 and its receptor: from signal transduction to tumorigenesis. Growth Factors. 2004; 22:207-215. (Biology). View Reference
    6. Merz H, Houssiau FA, Orscheschek K, et al. Interleukin-9 expression in human malignant lymphomas: unique association with Hodgkin's disease and large cell anaplastic lymphoma. Blood. 1991 September; 78(5):1311-1317. (Biology). View Reference
    7. Renauld JC. New insights into the role of cytokines in asthma. J Clin Pathol. 2001 August; 54(8):577-589. (Biology). View Reference
    8. Soler D, Chapman TR, Poisson LR. CCR8 expression identifies CD4 memory T cells enriched for FOXP3+ regulatory and Th2 effector lymphocytes. J Immunol. 2006; 177(10):6940-6951. (Biology). View Reference
    9. Soroosh P, Doherty TA. Th9 and allergic disease. Immunology. 2009; 127(4):450-458. (Biology). View Reference
    View All (9) View Less
    564254 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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