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      Alexa Fluor™ 488 Rat Anti-Human IL-10
      Alexa Fluor™ 488 Rat Anti-Human IL-10
      Multiparameter flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. An enriched population of CD4+ peripheral blood mononuclear cells (PBMC) was obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody (Cat. No. 555343). The CD4+ 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; 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) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). After harvest the cells were fixed using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained in BD Perm/Wash™ Buffer with BD Horizon™ BV421 Mouse Anti- Human CD4 antibody (Cat. No. 565997/566907) and with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human IL-10 antibody (Cat. No. 567010/567011; Right Plot). A bivariate pseudocolor density plot showing the coexpressed levels 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™ Cell Analyzer System and FlowJo™ software.
      Alexa Fluor™ 488 Rat Anti-Human IL-10
      Multiparameter flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. An enriched population of CD4+ peripheral blood mononuclear cells (PBMC) was obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody (Cat. No. 555343). The CD4+ 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; 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) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724). After harvest the cells were fixed using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained in BD Perm/Wash™ Buffer with BD Horizon™ BV421 Mouse Anti- Human CD4 antibody (Cat. No. 565997/566907) and with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human IL-10 antibody (Cat. No. 567010/567011; Right Plot). A bivariate pseudocolor density plot showing the coexpressed levels 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™ Cell Analyzer System and FlowJo™ software.
      製品詳細
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      BD Pharmingen™
      Interleukin-10; CSIF; Cytokine synthesis inhibitory factor; TGIF
      Human (QC Testing), Viral (Tested in Development)
      Rat IgG2a, κ
      Recombinant Human IL-10
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      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 unreacted dye was removed.
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      推奨アッセイ手順

      BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. 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.
      5. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      567011 Rev. 1
      抗体の詳細
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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).

              

      567011 Rev. 1
      フォーマットの詳細
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      Alexa Fluor™ 488
      Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 488
      Blue 488 nm
      494 nm
      517 nm
      567011 Rev.1
      引用＆参考文献
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      Development References (8)

      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. 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
      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. (Clone-specific: Bioassay, Functional assay, Inhibition). View Reference
      6. Wehrens EJ, Mijnheer G, Duurland CL, et al. Functional human regulatory T cells fail to control autoimmune inflammation due to PKB/c-akt hyperactivation in effector cells.. Blood. 2011; 118(13):3538-48. (Clone-specific: Flow cytometry). View Reference
      7. 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
      8. 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
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      567011 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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