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      APC Rat Anti-Mouse IL-4
      APC Rat Anti-Mouse IL-4
      Expression of IL-4 by stimulated CD4+ and CD4-Balb/c spleen cells. BALB/c spleen cells were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B (2 µg/ml final concentration; Sigma, St. Louis, MO), recombinant mouse IL-2 (10 U/ml final concentration; Cat. No. 550069) and recombinant mouse IL-4 (2 ng/ml final concentration; Cat. No. 550067). The cells were harvested and restimulated for 5 hours with anti-CD3 (2 µg/ml final concentration; 145-2C11, Cat. No. 553057) and anti-CD28 (2 µg/ml final concentration; clone 37.51, Cat. No. 553294) antibodies in the presence of GolgiStop™ (3 µM final concentration; Cat. No. 554724). The splenocytes were then stained with 0.25 µg of FITC-conjugated rat anti-mouse CD4 (FITC-RM4-5, Cat. No. 553047) and 0.25 µg of APC-conjugated rat anti-mouse IL-4 antibody (APC-11B11, Cat. No. 554436) by using Pharmingen's staining protocol (left panel). To demonstrate staining specificity, the binding of APC-11B11 was blocked by the preincubation of the conjugated antibody with excess recombinant mouse IL-4 (0.25 µg; Cat. No. 550037) (middle panel) or by pre-blocking fixed/permeabilized cells with excess purified \"cold\" 11B11 mouse antibody (5.0 µg; Cat. No. 554433) (right panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified using the cytokine-blocking and \"cold\" mouse antibody blocking controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.
      APC Rat Anti-Mouse IL-4
      Expression of IL-4 by stimulated CD4+ and CD4-Balb/c spleen cells. BALB/c spleen cells were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B (2 µg/ml final concentration; Sigma, St. Louis, MO), recombinant mouse IL-2 (10 U/ml final concentration; Cat. No. 550069) and recombinant mouse IL-4 (2 ng/ml final concentration; Cat. No. 550067). The cells were harvested and restimulated for 5 hours with anti-CD3 (2 µg/ml final concentration; 145-2C11, Cat. No. 553057) and anti-CD28 (2 µg/ml final concentration; clone 37.51, Cat. No. 553294) antibodies in the presence of GolgiStop™ (3 µM final concentration; Cat. No. 554724). The splenocytes were then stained with 0.25 µg of FITC-conjugated rat anti-mouse CD4 (FITC-RM4-5, Cat. No. 553047) and 0.25 µg of APC-conjugated rat anti-mouse IL-4 antibody (APC-11B11, Cat. No. 554436) by using Pharmingen's staining protocol (left panel). To demonstrate staining specificity, the binding of APC-11B11 was blocked by the preincubation of the conjugated antibody with excess recombinant mouse IL-4 (0.25 µg; Cat. No. 550037) (middle panel) or by pre-blocking fixed/permeabilized cells with excess purified \"cold\" 11B11 mouse antibody (5.0 µg; Cat. No. 554433) (right panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified using the cytokine-blocking and \"cold\" mouse antibody blocking controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.
      Product Details
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      BD Pharmingen™
      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_398556
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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

      Immunofluorescent Staining and Flow Cytometric Analysis: The APC- conjugated 11B11 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-4 producing cells within mixed cell populations. The use of an isotype control, such as rat IgG1 isotype control, APC-R3-34 (Cat. 554686) is recommended. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the Technical Resources/protocols section or the chapter on intracellular staining.

      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      554436 Rev. 3
      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.

      554436 Rev. 3
      Format Details
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      APC
      Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      APC
      Red 627-640 nm
      651 nm
      660 nm
      554436 Rev.3
      Citations & References
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      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
      554436 Rev. 3

      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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