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    BV421 Mouse Anti-Human IL-8
    BV421 Mouse Anti-Human IL-8
    Two-color flow cytometric analysis of IL-8 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 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Panel) or BD Horizon™ BV421 Anti-Human IL-8 antibody (Cat. No. 563310; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-8 (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 Mouse Anti-Human IL-8
    Two-color flow cytometric analysis of IL-8 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 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Panel) or BD Horizon™ BV421 Anti-Human IL-8 antibody (Cat. No. 563310; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-8 (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™
    IL8; CXCL8; GCP-1; LYNAP; MDNCF; MONAP; NAP-1; emoctakin
    Human (QC Testing)
    Mouse IgG2b
    Recombinant Human IL-8
    Intracellular staining (flow cytometry) (Routinely Tested)
    5 µl
    AB_2738131
    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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    563310 Rev. 1
    Antibody Details
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    G265-8

    The G265-8 monoclonal antibody specifically binds to both the 72 and 77 amino acid isoforms of human Interleukin-8 (IL-8). IL-8 is secreted as an 8-9 kDa, non-glycosylated proinflammatory chemokine protein also known as chemokine (C-X-C motif) ligand 8 (CXCL8). IL-8 is synthesized as a 99 amino acid precursor that is proteolytically processed into several isoforms. The 72 amino acid isoform is produced by monocytes, macrophages, granulocytes, epithelial cells, and fibroblasts in response to pro-inflammatory stimuli including cytokines and microbial agents. It is also expressed by endothelial cells, fibroblasts, keratinocytes, lymphocytes, and a variety of tumor cells. In response to IL-4, IL-10 and TGFβ, the cellular production of IL-8 is inhibited. IL-8 is crucial for the activation and recruitment of neutrophils to inflammatory sites. IL-8 is also a chemoattractant for basophils and T-lymphocytes. IL-8 possesses angiogenic activity and can be associated with tumor angiogenesis and metastasis. The 77 amino acid IL-8 isoform is primarily produced by endothelial cells. This larger isoform is reportedly a less potent neutrophil activator than the 72 amino acid isoform. IL-8 binds to and signals through two G-protein-coupled receptors, IL-8RA (CXCR1/CD181) and IL-8RB (CXCR2/CD182).

    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.

    563310 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
    563310 Rev.1
    Citations & References
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    Development References (9)

    1. Car BD, Meloni F, Luisetti M, Semenzato G, Gialdroni-Grassi G, Walz A. Elevated IL-8 and MCP-1 in the bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis and pulmonary sarcoidosis. Am J Respir Crit Care Med. 1994; 149(3 Pt 1):655-659. (Biology). View Reference
    2. Emadi S, Clay D, Desterke C, et al. IL-8 and its CXCR1 and CXCR2 receptors participate in the control of megakaryocytic proliferation, differentiation, and ploidy in myeloid metaplasia with myelofibrosis. Blood. 2005; 105(2):464-473. (Clone-specific: Flow cytometry, Immunocytochemistry (cytospins), Immunofluorescence). View Reference
    3. Hebert CA, Luscinskas FW, Kiely JM, et al. Endothelial and leukocyte forms of IL-8. Conversion by thrombin and interactions with neutrophils. J Immunol. 1990; 145(9):3033-3040. (Biology). View Reference
    4. Larsen CG, Anderson AO, Appella E, Oppenheim JJ, Matsushima K. The neutrophil-activating protein (NAP-1) is also chemotactic for T lymphocytes. Science. 1989; 243(4897):1464-1466. (Biology). View Reference
    5. Leonard EJ, Skeel A, Yoshimura T, Noer K, Kutvirt S, Van Epps D. Leukocyte specificity and binding of human neutrophil attractant/activation protein-1. J Immunol. 1990; 144(41323):1323-1330. (Biology). View Reference
    6. Lizasa H, Matsushima K. IL-8. 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; 2001:1061-1067.
    7. Matsushima K, Oppenheim JJ. Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine. 1989; 1(1):2-13. (Biology). View Reference
    8. Miller LS, Sorensen OE, Liu PT, et al. TGF-alpha regulates TLR expression and function on epidermal keratinocytes. J Immunol. 2005; 174(10):6137-6143. (Clone-specific: ELISA). View Reference
    9. 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. (Biology). View Reference
    View All (9) View Less
    563310 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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