Western blot using EGFR polyclonal antibody (Cat # PAB10223) shows detection of a band at ~170 KDa corresponding to human EGFR present in unstimulated (Lane 1) and EGF (50 ng/mL for 15 min) stimulated (Lane 2) A-431 whole cell lysates (arrowhead). Approximately 30 ug of lysate was resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:1,000. Reaction occurred overnight at 4°C followed by washes and reaction with a 1:10,000 dilution of IRDye® 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc.
Immunohistochemical staining with EGFR polyclonal antibody (Cat # PAB10223) was diluted to 2.5 ug/mL detect EGFR in human ovarian cancer tissue. Tissue was formalin fixed and paraffin embedded. No pre-treatment of sample was required. The image shows the localization of antibody as the precipitated red signal, with a hematoxylin purple nuclear counter stain.
Immunoprecipitation
Combined immunoprecipitation and immunoblot using EGFR polyclonal antibody (Cat # PAB10223). Lysates were prepared from GN4 rat liver epithelial cells both with (+) EGF treatment for 15' at 100 ng/mL and without (-) the addition of EGF. The combination of immunoprecipitation and immunoblotting was performed using the EGFR polyclonal antibody (Cat # PAB10223) for immunoprecipitation (10 uL) followed by immunoblot detection using an anti-phosphotyrosine antibody (Panel A). This was repeated in reverse order using a 1:2000 dilution of EGFR polyclonal antibody for immunoblot (Panel B). Visualization occurred using an ECL system. Film exposure was approximately 1'.
The protein encoded by this gene is a transmembrane glycoprotein that is a member of the protein kinase superfamily. This protein is a receptor for members of the epidermal growth factor family. EGFR is a cell surface protein that binds to epidermal growth factor. Binding of the protein to a ligand induces receptor dimerization and tyrosine autophosphorylation and leads to cell proliferation. Mutations in this gene are associated with lung cancer. [provided by RefSeq
Other Designations:
avian erythroblastic leukemia viral (v-erb-b) oncogene homolog,cell growth inhibiting protein 40,cell proliferation-inducing protein 61,epidermal growth factor receptor