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      PE-Cy7 Rat Anti-Human IL-10
      PE-Cy7 Rat Anti-Human IL-10
      Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody (Cat. No. 555343). The cells 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; 20 ng/ml) and IL-4 (Cat. No. 554605; 40 ng/ml) proteins. The cells were restimulated (5 h) with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P8139; 50 ng/ml) and Ionomycin (Sigma I9657; 1 μg/ml) and BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The cells were harvested, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti- Human CD4 antibody (Cat. No. 565997/566907) and with either PE-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552784; Left Plot) or PE-Cy7 Mouse Anti-Human IL-10 antibody (Cat. No. 567407/567408; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of IL-10 (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
      PE-Cy7 Rat Anti-Human IL-10
      Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody (Cat. No. 555343). The cells 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; 20 ng/ml) and IL-4 (Cat. No. 554605; 40 ng/ml) proteins. The cells were restimulated (5 h) with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P8139; 50 ng/ml) and Ionomycin (Sigma I9657; 1 μg/ml) and BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). The cells were harvested, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti- Human CD4 antibody (Cat. No. 565997/566907) and with either PE-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552784; Left Plot) or PE-Cy7 Mouse Anti-Human IL-10 antibody (Cat. No. 567407/567408; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of IL-10 (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
      Product Details
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      BD Pharmingen™
      Interleukin-10; CSIF; Cytokine synthesis inhibitory factor; TGIF
      Human (QC Testing)
      Rat IgG2a, κ
      Recombinant Human IL-10
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl/test
      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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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).
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
      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. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
      8. An isotype control should be used at the same concentration as the antibody of interest.
      9. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
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      567407 Rev. 1
      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).

      567407 Rev. 1
      Format Details
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      PE-Cy7
      PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-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.
      altImg
      PE-Cy7
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      781 nm
      567407 Rev.1
      Citations & References
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      View product citations for antibody "567407" on CiteAb

      Development References (9)

      1. Andersson J, Abrams J, Bjork L, et al. Concomitant in vivo production of 19 different cytokines in human tonsils. Immunology. 1994; 83(1):16-24. (Clone-specific: Immunohistochemistry). View Reference
      2. Brodeur ND, Spencer JV. Antibodies to human IL-10 neutralize ebvIL-10-mediated cytokine suppression but have no effect on cmvIL-10 activity.. Virus Res. 2010; 153(2):265-8. (Clone-specific: Bioassay, Blocking, ELISA, Functional assay, Inhibition, Neutralization, Western blot). View Reference
      3. Cousins DJ, Lee TH, Staynov DZ. Cytokine coexpression during human Th1/Th2 cell differentiation: direct evidence for coordinated expression of Th2 cytokines.. J Immunol. 2002; 169(5):2498-506. (Clone-specific: Flow cytometry). View Reference
      4. 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. (Biology). View Reference
      5. Mielle J, Audo R, Hahne M, et al. IL-10 Producing B Cells Ability to Induce Regulatory T Cells Is Maintained in Rheumatoid Arthritis. Front Immunol. 2018; 9:961. (Clone-specific: Flow cytometry). View Reference
      6. Moore KW, de Waal Malefyt R, Coffman RL, O'Garra A. Interleukin-10 and the interleukin-10 receptor.. Annu Rev Immunol. 2001; 19:683-765. (Biology). View Reference
      7. 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. (Clone-specific: Bioassay, Functional assay, Inhibition). View Reference
      8. Yin Y, Mitson-Salazar A, Prussin C. Detection of Intracellular Cytokines by Flow Cytometry. Curr Protoc Immunol. 2015; 110:6.24.21-26.24.18. (Methodology: Flow cytometry). View Reference
      9. Zielinski CE, Mele F, Aschenbrenner D, et al. Pathogen-induced human TH17 cells produce IFN-gamma or IL-10 and are regulated by IL-1beta. Nature. 2012; 484(7395):514-518. (Clone-specific: Flow cytometry). View Reference
      View All (9) View Less
      567407 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.