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      BV650 Rat Anti-Mouse IL-4
      BV650 Rat Anti-Mouse IL-4
      Two-color flow cytometric analysis of IL-4 expression in stimulated mouse T cells. An enriched population of mouse splenic CD4+ T cells were cultured (2 days) with plate-bound Purified NA/L E Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml) antibodies with Recombinant IL-2 (Cat. No. 550069; 10 ng/ml) and IL-4 (Cat. No. 550067; 25 ng/ml). The cells were subsequently cultured (3 days) with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA (Sigma Cat. No. P-8139) and Ionomycin (Sigma Cat. No. I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) for 6 hours. After harvesting, the cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then washed, fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722).        The cells were subsequently washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BV650 Rat IgG1, κ Isotype Control (Cat. No. 563848; Left Panel) or BD Horizon BV650 Rat Anti-Mouse IL-4 antibody (Cat. No . 564004; Right Panel). Flow cytometric contour plots showing correlated expression of IL-4 (or Ig Isotype control staining) versus Autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      BV650 Rat Anti-Mouse IL-4
      Two-color flow cytometric analysis of IL-4 expression in stimulated mouse T cells. An enriched population of mouse splenic CD4+ T cells were cultured (2 days) with plate-bound Purified NA/L E Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml) antibodies with Recombinant IL-2 (Cat. No. 550069; 10 ng/ml) and IL-4 (Cat. No. 550067; 25 ng/ml). The cells were subsequently cultured (3 days) with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA (Sigma Cat. No. P-8139) and Ionomycin (Sigma Cat. No. I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) for 6 hours. After harvesting, the cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then washed, fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722).        The cells were subsequently washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BV650 Rat IgG1, κ Isotype Control (Cat. No. 563848; Left Panel) or BD Horizon BV650 Rat Anti-Mouse IL-4 antibody (Cat. No . 564004; Right Panel). Flow cytometric contour plots showing correlated expression of IL-4 (or Ig Isotype control staining) versus Autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      제품 세부 정보
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      BD Horizon™
      Ms IL-4 BV650 11B11 50ug
      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_2687569
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      준비 및 보관

      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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      권장 분석 절차

      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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      제품 고시

      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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      7. 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.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
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      관련 제품

      Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
      사이즈 0.1 mg 카탈로그 No 553141
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      Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
      사이즈 0.5 mg 카탈로그 No 553142
      sampleImage/
      Stain Buffer (FBS) RUO
      사이즈 500 mL 카탈로그 No 554656
      sampleImage/
      Fixation and Permeabilization Solution RUO
      사이즈 125 mL 카탈로그 No 554722
      sampleImage/
      Perm/Wash Buffer RUO
      사이즈 100 mL 카탈로그 No 554723
      sampleImage/
      Protein Transport Inhibitor (Containing Monensin) RUO
      사이즈 0.7 mL 카탈로그 No 554724
      sampleImage/
      세부 정보보기
      564004 Rev. 2
      항체 세부 정보
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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.

      564004 Rev. 2
      형광 세부 정보
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      BV650
      The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-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
      BV650
      Violet 405 nm
      406 nm
      649 nm
      564004 Rev.2
      인용 및 참고 문헌
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      개발 참고 자료 (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). 참조 보기
      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). 참조 보기
      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: Functional assay, Neutralization). 참조 보기
      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: Functional assay, Neutralization). 참조 보기
      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). 참조 보기
      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). 참조 보기
      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: Flow cytometry, IC/FCM Block). 참조 보기
      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: Functional assay, Neutralization). 참조 보기
      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). 참조 보기
      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: Functional assay, Neutralization). 참조 보기
      모두 보기 (10) 간단히 보기
      564004 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.

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