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Biotin Rat Anti-Mouse CD124
Biotin Rat Anti-Mouse CD124

Expression of cell surface CD124 by B220-positve and -negative splenic lymphocytes from C57BL/6 mice. Spleen cells from C57BL/6 mice were treated with ACK lysis buffer, washed, and were labeled with purified mouse BD Fc Block™ (Cat. No. 553142) to block mouse Fc receptors. The cells were then stained with biotinylated mIL4R-M1 (0.125 µg/10^6 cells) followed by streptavidin  phycoerythrin (0.015 µg, Cat. No. 554061) and FITC anti-mouse B220 (0.06 µg, Cat. No. 553088) and were analyzed by two-color flow cytometry. The levels of CD124 expressed by B220-positive and B220-negative cells (with the light-scattering characteristics of viable lymphocytes) are shown in the two-color dot plot (right panel). Staining with the biotinylated mIL4R-M1 antibody (right panel) is compared to staining derived with a biotinylated rat IgG2a immunoglobulin isotype control antibody (0.125 µg, Cat. No. 553928) that is shown in the left panel.

Expression of cell surface CD124 by B220-positve and -negative splenic lymphocytes from C57BL/6 mice. Spleen cells from C57BL/6 mice were treated with ACK lysis buffer, washed, and were labeled with purified mouse BD Fc Block™ (Cat. No. 553142) to block mouse Fc receptors. The cells were then stained with biotinylated mIL4R-M1 (0.125 µg/10^6 cells) followed by streptavidin  phycoerythrin (0.015 µg, Cat. No. 554061) and FITC anti-mouse B220 (0.06 µg, Cat. No. 553088) and were analyzed by two-color flow cytometry. The levels of CD124 expressed by B220-positive and B220-negative cells (with the light-scattering characteristics of viable lymphocytes) are shown in the two-color dot plot (right panel). Staining with the biotinylated mIL4R-M1 antibody (right panel) is compared to staining derived with a biotinylated rat IgG2a immunoglobulin isotype control antibody (0.125 µg, Cat. No. 553928) that is shown in the left panel.

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
ELISA (Routinely Tested), Flow cytometry, Neutralization (Tested During Development), Immunoprecipitation (Reported)
0.5 mg/ml
AB_394406
Aqueous buffered solution containing protein stabilizer 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 biotin under optimum conditions, and unreacted biotin was removed.

Recommended Assay Procedures

Immunofluorescent staining and Flow Cytometric Analysis: The biotinylated form of the mIL4R-M1 antibody can be used for the immunofluorescent staining and flow cytometric analysis of nucleated mouse cells to measure their expressed levels of surface CD124. An appropriate biotin conjugated Ig isotype control is clone R35-95 (Cat. No. 553928).

ELISA: The biotinylated mIL4R-M1 antibody can be paired with the mIL4R-M2 antibody, (Cat. No. 552952) for use in a sandwich ELISA for measuring soluble mouse CD124 protein levels  and with purified recombinant protein as the standard. The capture antibody should be titrated between 1-4 µg/ml to determine its optimal coating concentration. The mIL4R-M1 detection mAb should be titrated between 0.5-2 µg/ml.  To obtain linear standard curves, doubling dilutions of recombinant soluble mouse CD124  ranging from 2000 to 15 pg/ml are recommended for inclusion in each ELISA plate.

Neutralization: The mIL4R-M1 antibody blocks the binding of IL-4 to the IL-4 receptor. For use in this application our Cat. No. 552288, in NA/LE format 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.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
552508 Rev. 2
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.

552508 Rev. 2
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
552508 Rev.2
Citations & References
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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. (Biology: 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
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552508 Rev. 2

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