Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified mIL-4R-M2 antibody (Cat. No. 551853) is useful as a capture antibody for a sandwich ELISA that measures soluble mouse IL-4Rα protein levels. The purified mIL-4R-M2 antibody can be paired with biotinylated mIL-4R-M1 antimouse IL-4Rα antibody (Cat. No. 552508) as the detection antibody and with purified recombinant mouse IL-4Rα protein as the standard. This capture antibody should be titrated between 1-4 µg/ml to determine its optimal coating concentration for ELISA. To obtain linear standard curves, doubling dilutions of recombinant soluble mouse IL-4Rα ranging from 2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.
Note: This ELISA antibody pair shows no cross-reactivity with the following recombinant mouse proteins: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-7, IL-10, IL-13, IL-15, IL-18, TNF, sTNFRI, sTNFRII, GM-CSF, CSF-1, Granzyme B, and IFN-γ.
Immunoprecipitation: The mIL4R-M2 antibody is reported to immunoprecipitate mouse IL-4Rα proteins. Please note that this application is not routinely tested at BD Biosciences.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
The mIL4R-M2 monoclonal antibody specifically binds to the α subunit of the mouse Interleukin-4 Receptor (IL-4Rα, aka, CD124). The mIL4R-M2 antibody does not recognize human or rat IL-4R; crossreactivities with IL-4R from other species have not been established. The mouse IL-4Rα is a 140 kDa transmembrane glycoprotein that is expressed by B and T lymphocytes and a variety of other hematopoietic and nonhematopoietic cells and cell lines. The cell surface IL-4Rα chain binds IL-4 with high affinity and associates with either the common γ chain (IL-4Rα/γc; aka, type I IL-4R) or the IL-13 receptor alpha subunit (IL-4Rα/IL-13Rα; aka, type II IL-4R complex) to form two distinct types of signal-transducing IL-4R complexes. The type I IL-4 receptor complex specifically binds IL-4 whereas the type II IL-4R binds and transduces signals from either IL-4 or IL-13. The mIL4R-M2 antibody blocks IL-4 binding to cells and is reported to be a potent inhibitor of IL-4's biological activities. The mIL4R-M2 antibody also recognizes naturally occurring, soluble truncated forms of IL-4Rα (sIL-4R) that result either from enzymatic cleavage of the cell surface extracellular IL-4Rα domain or from differential mRNA splicing and secretion by cells. These sIL-4R retain their high-affinity ligand binding domain and appear to either enhance or inhibit IL-4-mediated functions depending on the relative local levels of IL-4 and sIL-4R. The immunogen used to generate the mIL4R-M2 hybridoma was CTLL-19.4 cells, a subclone of CTLL-2 cells that express high levels of IL-4Rα.
Development References (8)
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Beckmann MP, Schooley KA, Gallis B, et al. Monoclonal antibodies block murine IL-4 receptor function. J Immunol. 1990; 144(11):4212-4217. (Immunogen: Blocking, Immunoprecipitation, Neutralization). View Reference
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Chilton PM, Fernandez-Botran R. Production of soluble IL-4 receptors by murine spleen cells is regulated by T cell activation and IL-4. J Immunol. 1993; 151(1):5907-5917. (Biology). View Reference
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Feldman GM, Ruhl S, Bickel M, Finbloom DS, Pluznik DH. Regulation of interleukin-4 receptors on murine myeloid progenitor cells by interleukin-6. Blood. 1991; 78(7):1678-1684. (Biology). View Reference
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Hassuneh MR, Nagarkatti PS, Nagarkatti M. Evidence for the participation of interleukin-2 (IL-2) and IL-4 in the regulation of autonomous growth and tumorigenesis of transformed cells of lymphoid origin. Blood. 1997; 89(2):610-620. (Biology). View Reference
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Kubo M, Yamashita M, Abe R, et al. CD28 costimulation accelerates IL-4 receptor sensitivity and IL-4-mediated Th2 differentiation. J Immunol. 1999; 63(5):2432-2442. (Biology). View Reference
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Lowenthal JW, Castle BE, Christiansen J, et al. Expression of high affinity receptors for murine interleukin 4 (BSF-1) on hemopoietic and nonhemopoietic cells. J Immunol. 1988; 140(2):456-464. (Biology). View Reference
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Mosley B, Beckmann MP, March CJ, et al. The murine interleukin-4 receptor: molecular cloning and characterization of secreted and membrane bound forms. Cell. 1989; 59(2):335-348. (Biology). View Reference
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Sempowski GD, Beckmann MP, Derdak S, Phipps RP. Subsets of murine lung fibroblasts express membrane-bound and soluble IL-4 receptors. Role of IL-4 in enhancing fibroblast proliferation and collagen synthesis. J Immunol. 1994; 152(7):3606-3614. (Biology). View Reference
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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.
