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      BV421 Rat Anti-Mouse IL-4
      BV421 Rat Anti-Mouse IL-4
      Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes.  Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates (GK1.5, Cat. No.553726;10 μg/ml) for 1 hr at 4°C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (145-2C11, Cat. No. 553057;10 μg/ml) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (37.51, Cat. No. 553294; 2 μg/ml) antibody and Recombinant Mouse IL-2 (Cat. No. 550069; 10 ng/ml) and IL-4 (Cat. No. 550067; 50 ng/ml) for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated (4 hr)  with PMA (Sigma, Cat. No. P-8139; 5 ng/ml) and ionomycin (Sigma, Cat. No. P-8139; 500 ng) in the presence of BD GolgiPlug™ Protein Transport Inhibitor (Containing Brefeldin A) (Cat. No. 555029).  The activated cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized using the BD Perm/Wash™ Permeabilization Buffer (Cat. No. 554723) and then stained either with a BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868, Left Panel) or with BD Horizon™ BV421 Rat Anti-Mouse IL-4 antibody (Cat. No. 562915, Right Panel). Two-color dot plots showing IL-4 (or Ig isotype control staining) versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR™ II Flow Cytometry System.
      BV421 Rat Anti-Mouse IL-4
      Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes.  Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates (GK1.5, Cat. No.553726;10 μg/ml) for 1 hr at 4°C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (145-2C11, Cat. No. 553057;10 μg/ml) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (37.51, Cat. No. 553294; 2 μg/ml) antibody and Recombinant Mouse IL-2 (Cat. No. 550069; 10 ng/ml) and IL-4 (Cat. No. 550067; 50 ng/ml) for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated (4 hr)  with PMA (Sigma, Cat. No. P-8139; 5 ng/ml) and ionomycin (Sigma, Cat. No. P-8139; 500 ng) in the presence of BD GolgiPlug™ Protein Transport Inhibitor (Containing Brefeldin A) (Cat. No. 555029).  The activated cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized using the BD Perm/Wash™ Permeabilization Buffer (Cat. No. 554723) and then stained either with a BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868, Left Panel) or with BD Horizon™ BV421 Rat Anti-Mouse IL-4 antibody (Cat. No. 562915, Right Panel). Two-color dot plots showing IL-4 (or Ig isotype control staining) versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR™ II Flow Cytometry System.
      Product Details
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      BD Horizon™
      IL4: Interleukin-4; BSF-1; B-cell growth factor 1; BCGF-1
      Mouse (QC Testing)
      Rat IgG1, κ
      Partially Purified Mouse IL-4
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      16189
      AB_2737889
      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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
      4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
      9. 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.
      10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      562915 Rev. 2
      Antibody Details
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      11B11

      Interleukin-4 (IL-4) is a pleiotropic cytokine that has many roles, such as inducing the differentiation of naïve helper T cells (Th0 cells) to Th2 cells, stimulating activated B-cell and T-cell proliferation, and promoting immunoglobulin class switching to IgG1 and IgE in mouse B-cells.  IL-4 is expressed by CD4 T-cells, mast cells, basophils and eosinophils.  IL-4 was previously known as B-Cell Differentiation Factor (BCDF) or B-cell Stimulatory Factor (BSF1).  The 11B11 monoclonal antibody specifically binds to mouse IL-4. The immunogen used to generate the 11B11 hybridoma was partially purified mouse IL-4 prepared from the supernatant of Phorbol 12-Myristate 13-Acetate (PMA)-stimulated EL-4 cells. The 11B11 antibody is reportedly a neutralizing antibody.

      562915 Rev. 2
      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
      562915 Rev.2
      Citations & References
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      View product citations for antibody "562915" on CiteAb

      Development References (10)

      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. Assenmacher M, Schmitz J, Radbruch A. Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 in interferon-gamma and in interleukin-4-expressing cells. Eur J Immunol. 1994; 24(5):1097-1101. (Clone-specific: Flow cytometry). View Reference
      3. Haak-Frendscho M, Brown JF, Iizawa Y, Wagner RD, Czuprynski CJ. Administration of anti-IL-4 monoclonal antibody 11B11 increases the resistance of mice to Listeria monocytogenes infection. J Immunol. 1992; 148(12):3978-3985. (Clone-specific: Neutralization). View Reference
      4. Lindqvist C, Lundstrom H, Oker-Blom C, Akerman KE. Enhanced IL-4-mediated D10.G4.1 proliferation with suboptimal concentrations of anti-IL-4 receptor monoclonal antibodies. J Immunol. 1993; 150(2):394-398. (Clone-specific: Neutralization). View Reference
      5. Ohara J, Paul WE. Production of a monoclonal antibody to and molecular characterization of B-cell stimulatory factor-1. Nature. 1985; 315(6017):333-336. (Immunogen). View Reference
      6. Openshaw P, Murphy EE, Hosken NA, et al. Heterogeneity of intracellular cytokine synthesis at the single-cell level in polarized T helper 1 and T helper 2 populations. J Exp Med. 1995; 182(5):1357-1367. (Clone-specific: Flow cytometry). View Reference
      7. 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). View Reference
      8. Sadick MD, Heinzel FP, Holaday BJ, Pu RT, Dawkins RS, Locksley RM. Cure of murine leishmaniasis with anti-interleukin 4 monoclonal antibody. Evidence for a T cell-dependent, interferon gamma-independent mechanism. J Exp Med. 1990; 171(1):115-127. (Clone-specific: Neutralization). View Reference
      9. Sander B, Hoiden I, Andersson U, Moller E, Abrams JS. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods. 1993; 166(2):201-214. (Clone-specific: ELISA, Flow cytometry). View Reference
      10. Swain SL, Weinberg AD, English M, Huston G. IL-4 directs the development of Th2-like helper effectors. J Immunol. 1990; 145(11):3796-3806. (Clone-specific: Neutralization). View Reference
      View All (10) View Less
      562915 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.