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      BB700 Rat Anti-Human IL-2
      BB700 Rat Anti-Human IL-2
      Two color flow cytometric analysis of IL-2 expression in activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml) and Calcium Ionophore A23187 (Sigma C-9275; 1 μg/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat No. 562426/562427) and either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat No. 566413; Left Plot) or BD Horizon BB700 Rat Anti-Human IL-2 antibody (Cat No. 566405/566406; Right Plot) using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric contour plots showing the correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
      BB700 Rat Anti-Human IL-2
      Two color flow cytometric analysis of IL-2 expression in activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml) and Calcium Ionophore A23187 (Sigma C-9275; 1 μg/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat No. 562426/562427) and either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat No. 566413; Left Plot) or BD Horizon BB700 Rat Anti-Human IL-2 antibody (Cat No. 566405/566406; Right Plot) using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric contour plots showing the correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
      Product Details
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      BD Horizon™
      IL2; Interleukin-2; T-cell growth factor; TCGF
      Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
      Rat IgG2a, κ
      Human IL-2 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      3558
      AB_2744488
      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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).

      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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      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. An isotype control should be used at the same concentration as the antibody of interest.
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      566405 Rev. 1
      Antibody Details
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      MQ1-17H12

      The MQ1-17H12 monoclonal antibody specifically binds to the multifunctional cytokine, human Interleukin-2 (IL-2). IL-2 is produced by activated T cells and has multiple functions that can affect the growth, proliferation, differentiation and survival of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the MQ1-17H12 hybridoma was purified recombinant human IL-2 protein. The MQ1-17H12 antibody reportedly neutralizes the biological activity of human IL-2.

      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.

      566405 Rev. 1
      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
      566405 Rev.1
      Citations & References
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      View product citations for antibody "566405" on CiteAb

      Development References (6)

      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). 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: Blocking, ELISA, Immunoprecipitation). View Reference
      3. Mascher B, Schlenke P, Seyfarth M. Expression and kinetics of cytokines determined by intracellular staining using flow cytometry. J Immunol Methods. 1999; 223(1):115-121. (Clone-specific: Flow cytometry). 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. Smith, KA. Interleukin-2: inception, impact, and implications. Science. 1988; 240(4856):1169-1176. (Biology). View Reference
      6. Taniguchi T, Matsui H, Fujita T, et al. Structure and expression of a cloned cDNA for human interleukin-2. Nature. 1983; 302(5906):305-310. (Biology). View Reference
      View All (6) View Less
      566405 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.