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      Purified Rat Anti-Mouse CD124
      Purified Rat Anti-Mouse CD124
      Expression of cell surface CD124 by normal splenocyres from BALB/c and C57BL/6 mice. Spleen cells from BALB/c (left panel) and C57BL/6 (right panel) mice were treated with ACK lysis buffer, were washed, and were labeled with purified Fc Block (Cat. No. 553142; Rat IgG2b anti-mouse CD16/CD32) to block mouse Fc receptors. The cells were then stained with mIL4R-M1 (1 µg) followed by biotinylated RG7/1.30 (mouse anti-rat IgG2α; 0.25 µg, Cat. No. 553894) and streptavidin phycoerythrin (0.015 µg, Cat. No. 554061) in a three-layer staining protocol to amplify immunofluorescent signals. Staining with the mIL4R-M1 antibody (filled histograms) is compared to staining with the secondary reagents alone (open histograms).
      Purified Rat Anti-Mouse CD124
      Expression of cell surface CD124 by normal splenocyres from BALB/c and C57BL/6 mice. Spleen cells from BALB/c (left panel) and C57BL/6 (right panel) mice were treated with ACK lysis buffer, were washed, and were labeled with purified Fc Block (Cat. No. 553142; Rat IgG2b anti-mouse CD16/CD32) to block mouse Fc receptors. The cells were then stained with mIL4R-M1 (1 µg) followed by biotinylated RG7/1.30 (mouse anti-rat IgG2α; 0.25 µg, Cat. No. 553894) and streptavidin phycoerythrin (0.015 µg, Cat. No. 554061) in a three-layer staining protocol to amplify immunofluorescent signals. Staining with the mIL4R-M1 antibody (filled histograms) is compared to staining with the secondary reagents alone (open histograms).
      Product Details
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      BD Pharmingen™
      Il4r; IL-4Rα; Il4ra; IL-4RA; IL-4R-alpha; IL4-BP; IL-4 Receptor α chain
      Mouse (QC Testing)
      Rat LEW, also known as Lewis IgG2a, κ
      Mouse CTLL-19.4 T Cell Line
      Flow cytometry (Routinely Tested), ELISA (Tested During Development), Immunoprecipitation, Neutralization (Reported)
      0.5 mg/ml
      AB_394274
      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.
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      Recommended Assay Procedures

      Immunofluorescent Staining and Flow Cytometric Analysis: The purified form of the mIL4R-M1 antibody can be used for the immunofluorescent staining (≤ 1 µg antibody/10e6 cells) and flow cytometric analysis of nucleated mouse cells to measure their expressed levels of surface CD124. An appropriate purified Ig isotype control is clone R35-95 (Cat. No. 553927). A three-layer staining protocol is recommended for maximizing the detection of IL-4Rα expressed by cells as detailed in the figure legend.

      ELISA: The purified mIL4R-M1 antibody is useful for a sandwich ELISA that measures soluble mouse CD124 protein levels. The biotinylated mIL-4R-M1 antibody (Cat. No. 552508) can be paired with the purified mIL4R-M2 (rat anti-mouse sIL-4R) antibody, Cat. No. 552952, and recombinant soluble mIL-4Rα as the standard.

      Neutralization: The mIL4R-M1 antibody reportedly blocks IL-4's binding to and biological effects on IL-4R-positive cells. Note: for use in in vivo and in vitro systems, our NA/LE (no azide, low endotoxin) format, Cat. No. 552288 is recommended.

      Immunoprecipitation: The mIL4R-M1 antibody is reported to immunoprecipitate mouse IL-4R proteins. Please note that this application is not routinely tested at BD Biosciences Pharmingen.

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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. 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.
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      551853 Rev. 1
      Antibody Details
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      mIL4R-M1

      The mIL4R-M1 monoclonal antibody specifically binds to CD124 which is also known as the α subunit of the mouse Interleukin-4 Receptor (IL-4Rα). 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-M1 antibody blocks IL-4 binding to cells and is reported to be a potent inhibitor of IL-4's biological activities. The mIL4R-M1 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 mRNAsplicing 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.

      551853 Rev. 1
      Format Details
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      Purified
      Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
      Purified
      551853 Rev.1
      Citations & References
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      View product citations for antibody "551853" on CiteAb

      Development References (9)

      1. Beckmann MP, Schooley KA, Gallis B, et al. Monoclonal antibodies block murine IL-4 receptor function. J Immunol. 1990; 144(11):4212-4217. (Clone-specific: Immunoprecipitation, Neutralization). View Reference
      2. 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. (Clone-specific: ELISA). View Reference
      3. 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. (Clone-specific: Flow cytometry). View Reference
      4. Gessner A, Rollinghoff M. Biologic functions and signaling of the interleukin-4 receptor complexes. Immunobiology. 2000; 201(3-4):285-307. (Biology). View Reference
      5. 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. (Clone-specific: Flow cytometry). View Reference
      6. 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. (Clone-specific: Flow cytometry). View Reference
      7. 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
      8. 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
      9. 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. (Clone-specific: Flow cytometry). View Reference
      View All (9) View Less
      551853 Rev. 1

       

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      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.