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    BV421 Rat Anti-Human IL-6
    BV421 Rat Anti-Human IL-6
    Two-color flow cytometric analysis of IL-6 expression in stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were primed for 2 hr with Recombinant Human IFN-γ (10 ng/ml; Cat. No. 554617/554616) and stimulated with lipopolysaccharide (1.0 µg/ml; Sigma, Cat. No. L-8274; 6 hr) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing monensin) (Cat. No. 554724). The PBMC were then stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383). After washing with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), the cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then stained with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; Left Panel) or BD Horizon™ BV421 Anti-Human IL-6 antibody (Cat. No. 563279; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-6 (or Ig Isotype control staining) versus CD14 for gated events with the forward and side light-scatter characteristics of intact stimulated PBMC. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
    BV421 Rat Anti-Human IL-6
    Two-color flow cytometric analysis of IL-6 expression in stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were primed for 2 hr with Recombinant Human IFN-γ (10 ng/ml; Cat. No. 554617/554616) and stimulated with lipopolysaccharide (1.0 µg/ml; Sigma, Cat. No. L-8274; 6 hr) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing monensin) (Cat. No. 554724). The PBMC were then stained with APC Mouse Anti-Human CD14 antibody (Cat. No. 555399/561708/561383). After washing with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), the cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then stained with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; Left Panel) or BD Horizon™ BV421 Anti-Human IL-6 antibody (Cat. No. 563279; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-6 (or Ig Isotype control staining) versus CD14 for gated events with the forward and side light-scatter characteristics of intact stimulated PBMC. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
    Product Details
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    BD Horizon™
    IL6; Interleukin-6; BSF-2; CDF; HGF; HSF; IFNB2
    Human (QC Testing)
    Rat IgG1
    Human IL-6 Recombinant Protein
    Intracellular staining (flow cytometry) (Routinely Tested)
    5 µl
    AB_2738113
    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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
    3. An isotype control should be used at the same concentration as the antibody of interest.
    4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
    5. 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.
    6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
    8. Brilliant Violet™ 421 is a trademark of Sirigen.
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    563279 Rev. 1
    Antibody Details
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    MQ2-13A5

    The MQ2-13A5 monoclonal antibody specifically binds to human interleukin-6 (IL-6). IL-6 is a multifunctional cytokine that plays a central role in host defense mechanisms, including hematopoiesis, immune responses (eg, T and B cell activation and differentiation) and acute phase reactions. IL-6 can be expressed by a variety of cells including monocytes/macrophages, eosinophils, fibroblasts, vascular endothelial cells, bone marrow stromal cells, mesangial cells, hepatocytes, keratinocytes, astrocytes, T lymphocytes, B lymphocytes, and various tumor cells. IL-6 production is upregulated by cells in response to bacterial products such as lipopolysaccharide, viruses and other pro-inflammatory cytokines such as IL-1, TNF , and IFN-γ. IL-6 transcription is downregulated by cells in response to IL-4 and IL-10. The functional IL-6 Receptor (IL-6R) complex consists of two transmembrane glycoproteins, an 80-kDa low-affinity ligand-binding receptor subunit (IL-6Rα/CD126) and a 130 kDa (gp130/CD130) subunit that binds to IL-6-IL-6Rα to form the high-affinity signal transducing complex. Abnormal expression of IL-6 is related to the pathogenesis of many diseases including neoplastic (eg, multiple myeloma) and autoimmune diseases (eg, rheumatoid arthritis). The immunogen used to generate this hybridoma was COS-7 -expressed recombinant human IL-6.

    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.

    563279 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.
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    BV421
    Violet 405 nm
    407 nm
    423 nm
    563279 Rev.1
    Citations & References
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    Development References (7)

    1. Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA, Neutralization). View Reference
    2. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA, Neutralization). View Reference
    3. Gaines Das RE, Poole S. The international standard for interleukin-6. Evaluation in an international collaborative study. J Immunol Methods. 1993; 160(2):147-153. (Clone-specific: ELISA, Neutralization). View Reference
    4. Hirano T, Yasukawa K, Harada H, et al.. Complementary DNA for a novel human interleukin (BSF-2) that induces B lymphocytes to produce immunoglobulin. Nature. 1986; 324(6092):73-76. (Biology). View Reference
    5. Matsuda T, Hirano T. IL-6. In: Oppenheim JJ, Feldmann M, Durum SK, Hirano T, Vilcek J, Nicola NA, ed. Cytokine Reference : A compendium of cytokines and other mediators of host defense. San Diego: Academic Press; 2001:537-563.
    6. 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
    7. Rincon M. Interleukin-6: from an inflammatory marker to a target for inflammatory diseases. Trends Immunol. 2012; 33(11):571-577. (Biology). View Reference
    View All (7) View Less
    563279 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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