About InVivoMAb anti-mouse EGFR (domain I) The EMab-300 monoclonal antibody reacts with domain I of the mouse epidermal growth factor receptor (EGFR), also known as ERBB, ERBB1, and HER1. EGFR is a transmembrane glycoprotein and a member of the ErbB family of receptor protein tyrosine kinases (RTKs). The extracellular region of EGFR has four domains, wherein domains I and III contain the ligand-binding site. EGFR ligands include EGF, AREG, TGF-α, epiregulin (EREG), epigen, betacellulin, and heparin-binding EGF. Upon ligand binding, EGFR dimerizes, and its activation through the autophosphorylation of its cytoplasmic domain triggers several downstream signaling pathways, including RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLC-γ-PKC, and STATs, which help the cell respond accurately to extracellular signals. EGFR is expressed ubiquitously, and in various tissues, it plays critical roles in the developmental, physiological, and regenerative processes. EGFR’s aberrant activation is a common feature of various malignancies. Using monoclonal antibodies to target the EGFR extracellular region has shown promising results in anti-cancer research, and currently approved therapeutic antibodies such as cetuximab, panitumumab, nimotuzumab, and necitumumab bind to EGFR's domain III, which, together with domain I, is well known to facilitate EGF-EGFR binding. InVivoMAb anti-mouse EGFR (domain I) Specifications IsotypeRat IgG1, κ Recommended Isotype Control(s)InVivoMAb rat IgG1 isotype control, anti-horseradish peroxidase Recommended Dilution BufferInVivoPure pH 7.0 Dilution Buffer ImmunogenmEGFR over-expressing LN229 cells Reported ApplicationsFlow cytometry For details on in vivo applications, please contact technicalservice@bioxcell.com FormulationPBS, pH 7.0 Contains no stabilizers or preservatives Endotoxin<2EU/mg (<0.002EU/μg) Determined by LAL gel clotting assay Purity>95% Determined by SDS-PAGE Sterility0.2 μm filtered ProductionPurified from cell culture supernatant in an animal-free facility PurificationProtein G Molecular Weight150 kDa StorageThe antibody solution should be stored at the stock concentration at 4°C. Do not freeze. Application ReferencesInVivoMAb anti-mouse EGFR (domain I) (CLONE: EMab-300)Goto N, Suzuki H, Tanaka T, Ishikawa K, Ouchida T, Kaneko MK, Kato Y (2023). "EMab-300 Detects Mouse Epidermal Growth Factor Receptor-Expressing Cancer Cell Lines in Flow Cytometry" Antibodies (Basel) 12(3):42. PubMedEpidermal Growth Factor Receptor (EGFR) overexpression or its mutation mediates the sustaining proliferative signaling, which is an important hallmark of cancer. Human EGFR-targeting monoclonal antibody (mAb) therapy such as cetuximab has been approved for clinical use in patients with colorectal cancers and head and neck squamous cell carcinomas. A reliable preclinical mouse model is essential to further develop the mAb therapy against EGFR. Therefore, sensitive mAbs against mouse EGFR (mEGFR) should be established. In this study, we developed a specific and sensitive mAb for mEGFR using the Cell-Based Immunization and Screening (CBIS) method. The established anti-mEGFR mAb, EMab-300 (rat IgG1, kappa), reacted with mEGFR-overexpressed Chinese hamster ovary-K1 (CHO/mEGFR) and endogenously mEGFR-expressed cell lines, including NMuMG (a mouse mammary gland epithelial cell) and Lewis lung carcinoma cells, using flow cytometry. The kinetic analysis using flow cytometry indicated that the KD of EMab-300 for CHO/mEGFR and NMuMG was 4.3 × 10-8 M and 1.9 × 10-8 M, respectively. These results indicated that EMab-300 applies to the detection of mEGFR using flow cytometry and may be useful to obtain the proof of concept in preclinical studies.
