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      BB700 Rat Anti-Human IL-10
      BB700 Rat Anti-Human IL-10
      Two-color flow cytometric analysis of IL-10 expression by stimulated CD14+ human monocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 24 hours with lipopolysaccharide (LPS; Sigma-Aldrich Cat. No. L3137; 1 µg LPS/ml final concentration) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The stimulated cells were harvested and stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397/561712/557153) and then fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat. No. 566413; Left Plot) or BD Horizon BB700 Rat Anti-Human IL-10 antibody (Cat. No. 566567/566568; Right Plot) following the BD Biosciences staining protocol. Two-color flow cytometric dot plots showing the correlated expression of CD14 versus IL-10 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer System.
      BB700 Rat Anti-Human IL-10
      Two-color flow cytometric analysis of IL-10 expression by stimulated CD14+ human monocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 24 hours with lipopolysaccharide (LPS; Sigma-Aldrich Cat. No. L3137; 1 µg LPS/ml final concentration) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The stimulated cells were harvested and stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397/561712/557153) and then fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714). The cells were washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat. No. 566413; Left Plot) or BD Horizon BB700 Rat Anti-Human IL-10 antibody (Cat. No. 566567/566568; Right Plot) following the BD Biosciences staining protocol. Two-color flow cytometric dot plots showing the correlated expression of CD14 versus IL-10 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer System.
      Product Details
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      BD Horizon™
      Interleukin-10; CSIF; Cytokine synthesis inhibitory factor; TGIF
      Human (QC Testing), Viral (Tested in Development)
      Rat IgG2a, κ
      Recombinant Human IL-10
      Flow cytometry (Routinely Tested)
      5 µl
      AB_2869786
      Aqueous buffered solution containing ≤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™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.
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      Recommended Assay Procedures

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

      When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

      For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
      6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
      7. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
      8. Cy is a trademark of GE Healthcare.
      9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      566568 Rev. 2
      Antibody Details
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      JES3-19F1

      The JES3-19F1 monoclonal antibody specifically recognizes human Interleukin-10 (IL-10) that is encoded by IL10.  IL-10 is also known as Cytokine Synthesis Inhibitory Factor (CSIF), B cell-derived T cell growth factor (B-TCGF), and T-cell growth inhibitory factor (TGIF). The JES3-19F1 antibody crossreacts with ebvIL-10 protein, the Epstein-Barr viral IL-10 homolog (viral IL-10 or vIL-10) encoded by the BCRF1 gene. IL-10 is produced by a variety of cells such as some activated T cells and B cells including regulatory T cells (Treg) and B cells (Breg), monocytes and macrophages, dendritic cells (DC), keratinocytes, and mast cells. IL-10 is a multifunctional cytokine that can downregulate immune and proinflammatory responses. IL-10 can act to reduce expression of major histocompatibility complex class II antigens, costimulatory molecules, or proinflammatory cytokines including IL-1β, IL-2, IL-3,  IL-12, IFN-γ, TNF or GM-CSF expressed by activated monocytes, macrophages, dendritic cells (DC), natural killer (NK) cells, or T cells. IL-10 has been shown to play a role in chronic viral infections. IL-10 can also enhance B cell survival, proliferation, and differentiation to become antibody-producing cells. The JES3-19F1 antibody reportedly neutralizes the biological activity of human IL-10 and ebvIL-10. IL-10 mediates its biological activities by signaling through a heterotetrameric receptor complex composed of the type II cytokine receptor subunits CD210a (IL-10 Rα) and CD210b (IL-10 Rβ).

      The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

      566568 Rev. 2
      Format Details
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      BB700
      The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BB700
      Blue 488 nm
      476 nm
      695 nm
      566568 Rev.2
      Citations & References
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      View product citations for antibody "566568" on CiteAb

      Development References (5)

      1. 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, ELISPOT, Flow cytometry, Neutralization). View Reference
      2. Bacchetta R, Bigler M, Touraine JL, et al. High levels of interleukin 10 production in vivo are associated with tolerance in SCID patients transplanted with HLA mismatched hematopoietic stem cells. J Exp Med. 1994; 179(2):493-502. (Clone-specific: Neutralization). View Reference
      3. D'Andrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G. Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells. J Exp Med. 1993; 178(3):1041-1048. (Clone-specific: Neutralization). View Reference
      4. 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
      5. Schlaak JF, Schmitt E, Hüls C, Meyer zum Büschenfelde KH, Fleischer B. A sensitive and specific bioassay for the detection of human interleukin-10. J Immunol Methods. 1994; 168(1):49-54. (Biology). View Reference
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      566568 Rev. 2

      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.