BD Pharmingen™ Alexa Fluor™ 647 Rat Anti-Mouse IL-13
Clone W19-895 (RUO)
Multiparameter flow cytometric analysis of IL-13 expression in stimulated Mouse T cells. An enriched preparation of Mouse splenic CD4+ T cells was cultured (2 days) with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 567114/567115; 10 μg/ml for coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294/567110) antibodies with Recombinant Mouse IL-2 (Cat. No. 550069; 10 ng/ml) and Recombinant Mouse IL-4 (Cat. No. 550067; 25 ng/ml) proteins. The cells were subsequently cultured (3 days) with Recombinant IL-2 and IL-4 and 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 5.5 hours.
The cells were washed and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The fixed cells were washed and then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with PE-Cy7 Rat Anti-Mouse CD4 antibody (Cat. No. 552775) and with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse IL-13 antibody (Cat. No. 570080/570161; Right Plot) at 0.125 µg/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of IL-13 expression in stimulated Mouse T cells. An enriched preparation of Mouse splenic CD4+ T cells was cultured (2 days) with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 567114/567115; 10 μg/ml for coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294/567110) antibodies with Recombinant Mouse IL-2 (Cat. No. 550069; 10 ng/ml) and Recombinant Mouse IL-4 (Cat. No. 550067; 25 ng/ml) proteins. The cells were subsequently cultured (3 days) with Recombinant IL-2 and IL-4 and 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 5.5 hours.
The cells were washed and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The fixed cells were washed and then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with PE-Cy7 Rat Anti-Mouse CD4 antibody (Cat. No. 552775) and with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse IL-13 antibody (Cat. No. 570080/570161; Right Plot) at 0.125 µg/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Preparation And Storage
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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The W19-895 monoclonal antibody specifically recognizes mouse Interleukin-13 (IL-13), which is encoded by Il13 (interleukin 13). IL-13 is a multifunctional immunoregulatory cytokine that shares many functional properties with IL-4. It functions as a key regulator of immune responses against parasitic infections and as a mediator of inflammation involved in asthma and allergies. IL-13 is secreted by T helper 2 (Th2)-like cells, NK cells, Th1-like cells, CD8+ T cells, Innate lymphoid type 2 cells (ILC2), mast cells, macrophages, monocytes, basophils, and eosinophils. IL-13 promotes B-cell proliferation and induces immunoglobulin class switching to IgE and expression of CD23 (FcϵRII). It also upregulates expression of β2 integrin family members CD11b, CD11c, CD18, and induces MHC class II and CD23 expression on monocytes and macrophages. IL-13 directs the functions of these cells as well as endothelial cells, smooth muscle cells, fibroblasts, and epithelial cells by regulating their production of pro-inflammatory mediators including IL-1, IL-6, IL-12, TNF, and prostaglandins and reactive oxygen and nitrogen intermediates. IL-13 is the central mediator of allergic asthma in lungs where it regulates eosinophilic inflammation, mucus secretion, and airway hyperresponsiveness. IL-13 and IL-4 can bind with high affinity and signal intracellularly through the cell surface Type 2 IL-4 Receptor Complex comprised of IL-13 R alpha 1 (CD213a) and IL-4R alpha (CD124) chains. IL-13 also binds with high affinity to cell surface or soluble forms of the IL-13 R alpha 2 (CD213a2) which prevents IL-13 from signaling through the Type 2 IL-4 Receptor Complex and thus controls the bioavailability of IL-13.
Development References (10)
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Hershey GK. IL-13 receptors and signaling pathways: an evolving web.. J Allergy Clin Immunol. 2003; 111(4):677-90; quiz 691. (Biology). View Reference
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Iwaszko M, Biały S, Bogunia-Kubik K. Significance of Interleukin (IL)-4 and IL-13 in Inflammatory Arthritis.. Cells. 2021; 10(11):3000. (Biology). View Reference
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McKenzie A, Zurawski G. Measurement of IL-13. In: Coligan, Kruisbeek, Shevak, Strober, ed. Current Protocols in Immunology. New York: John Wiley & Sons; 1994:18-19.
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McKenzie AN, Culpepper JA, de Waal Malefyt R, et al. Interleukin 13, a T-cell-derived cytokine that regulates human monocyte and B-cell function.. Proc Natl Acad Sci U S A. 1993; 90(8):3735-9. (Biology). View Reference
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McKenzie GJ, Fallon PG, Emson CL, Grencis RK, McKenzie AN. Simultaneous disruption of interleukin (IL)-4 and IL-13 defines individual roles in T helper cell type 2-mediated responses.. J Exp Med. 1999; 189(10):1565-72. (Biology). View Reference
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Wynn TA. IL-13 effector functions.. Annu Rev Immunol. 2003; 21:425-56. (Biology). View Reference
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Zurawski G, de Vries JE.. Interleukin 13, an interleukin 4-like cytokine that acts on monocytes and B cells, but not on T cells. Immunol Today. 1994; 15(1):19-26. (Biology). View Reference
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