Skip to main content Skip to navigation
BV421 Rat Anti-Human IL-13
BV421 Rat Anti-Human IL-13

Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 µg/ml for plate coating) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 2 µg/ml) antibodies and Recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml) and Human IL-4 (Cat. No. 554605; 25 ng/ml) for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate (Sigma, Cat. No. P-8139; 50 ng/ml) and Ionomycin (Sigma, Cat. No. I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested and washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD4 antibody (Cat No. 561841/555349/561840) and either BD Horizon™ BV421 Rat IgG1 κ Isotype Control (Cat. No. 562868; Left Panel) or BD Horizon™ BV421 Mouse Anti-Human IL-13 antibody (Cat. No. 563580; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 (or Ig Isotype control staining) versus CD4 for gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 µg/ml for plate coating) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 2 µg/ml) antibodies and Recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml) and Human IL-4 (Cat. No. 554605; 25 ng/ml) for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate (Sigma, Cat. No. P-8139; 50 ng/ml) and Ionomycin (Sigma, Cat. No. I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested and washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD4 antibody (Cat No. 561841/555349/561840) and either BD Horizon™ BV421 Rat IgG1 κ Isotype Control (Cat. No. 562868; Left Panel) or BD Horizon™ BV421 Mouse Anti-Human IL-13 antibody (Cat. No. 563580; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 (or Ig Isotype control staining) versus CD4 for gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
Down Arrow Up Arrow


BD Horizon™
Interleukin-13; NC30; ALRH; BHR1; P600
Human (QC Testing)
Rat IgG1
Human Recombinant IL-13
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
3596
AB_2738290
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
563580 Rev. 1
Antibody Details
Down Arrow Up Arrow
JES10-5A2

The JES10-5A2 monoclonal antibody specifically binds to human interleukin-13, IL-13. IL-13 is produced by activated T cells, mast cells and NK cells. IL-13 regulates IgE production by B cells and can suppress the cytotoxic activity of macrophages and their production of inflammatory mediators. The immunogen used to produce the JES10-5A2 hybridoma was COS-expressed recombinant human IL-13. This is a neutralizing antibody.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes.  With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421  can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421  conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

563580 Rev. 1
Format Details
Down Arrow Up Arrow
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
563580 Rev.1
Citations & References
Down Arrow Up Arrow

Development References (7)

  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). View Reference
  2. 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
  3. Andersson U, Andersson J. Immunolabeling of cytokine-producing cells in tissues and in suspension. In: Fradelizie D, Emelie D, ed. Cytokine Producing Cells. Paris: Inserm; 1994:32-49.
  4. Litton M, Andersson J, Bjork L, Fehniger T, Ulfgren AK, Andersson U. Cytoplasmic cytokine staining in individual cells. In: Debets and Savelkoul, ed. Human Cytokine Protocols. Humana Press; 1996.
  5. McKenzie ANJ, Matthews DJ. IL-13. In: Oppenheim JJ, Feldmann M, Durum SK, Hirano T, Vilcek J, Nicola NA, ed. Cytokine Reference : A compendium of cytokines and other mediators of host defense. San Diego: Academic Press; 2001:203-211.
  6. 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
  7. Schaerli P, Willimann K, Lang AB, Lipp M, Loetscher P, Moser B. CXC chemokine receptor 5 expression defines follicular homing T cells with B cell helper function. J Exp Med. 2000; 192(11):1553-1562. (Clone-specific: Flow cytometry). View Reference
View All (7) View Less
563580 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.