Description
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.
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.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
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.
