Immunofluorescence analysis of EGFR was performed using 90% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with EGFR (R19/48MIX) Mouse Monoclonal Antibody (Product # AHR5062) at 1:100 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Yeast, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified extracellular and cytoplasmic domains of human recombinant EGFR protein.|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||1:250|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Epidermal Growth Factor Receptor (EGFR) is a 175 kDa transmembrane glycoprotein belonging to tyrosine kinase superfamily. It acts as a receptor for epidermal growth factor (EGF) family proteins. Binding of EGFR to its ligands causes autophosphorylation of tyrosine kinase followed by activation of signal transduction pathways connected to cell proliferation and differentiation. Tyrosine 1068 present within the cytoplasmic domain of the receptor is a major autophosphorylation site that allows binding of Grb2 and activation of the Ras-Raf-ERK1 and ERK2 signaling pathway.
IP-MS enrichment of EGFR (LFQ intensity): EGFR was enriched 1108-fold from A549 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and EGFR antibody (Part No. AHR5062). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Dual targeting of EGFR and ERBB2 pathways produces a synergistic effect on cancer cell proliferation and migration in vitro.
AHR5062 was used in western blot to analyze the production of a synergistic effect on cancer cell proliferation and migration in vitro due to dual targeting of ERBB2 and EGFR pathways
|Gray ME,Lee S,McDowell AL,Erskine M,Loh QT,Grice O,Argyle DJ,Bergkvist GT||Veterinary and comparative oncology (null:null)||2016|
|Not Applicable||Not Cited||
H19 non coding RNA-derived miR-675 enhances tumorigenesis and metastasis of breast cancer cells by downregulating c-Cbl and Cbl-b.
AHR5062 was used in western blot to characterize enhancement of tumorigenesis and metastasis of breast cancer cells by downregulating c-Cbl and Cbl-b by H19 non coding RNA-derived miR-675
|Vennin C,Spruyt N,Dahmani F,Julien S,Bertucci F,Finetti P,Chassat T,Bourette RP,Le Bourhis X,Adriaenssens E||Oncotarget (6:29209)||2015|
EGFR blockade enriches for lung cancer stem-like cells through Notch3-dependent signaling.
AHR5062 was used in immunoprecipitation and western blot to examine the effects of kinase inhibtors that target the epidermal growth factor receptor in lung cancer
|Arasada RR,Amann JM,Rahman MA,Huppert SS,Carbone DP||Cancer research (74:5572)||2014|
The soluble form of the tumor suppressor Lrig1 potently inhibits in vivo glioma growth irrespective of EGF receptor status.
AHR5062 was used in western blot to examine the therapeutic potential of the soluble form of Lrig for treating glioblastoma.
|Johansson M,Oudin A,Tiemann K,Bernard A,Golebiewska A,Keunen O,Fack F,Stieber D,Wang B,Hedman H,Niclou SP||Neuro-oncology (15:1200)||2013|
|Human||Not Cited||Sialyl Lewis X modification of the epidermal growth factor receptor regulates receptor function during airway epithelial wound repair.||Allahverdian S,Wang A,Singhera GK,Wong BW,Dorscheid DR||Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology (40:607)||2010|
An evolved Mxe GyrA intein for enhanced production of fusion proteins.
AHR5062 was used in flow cytometry to optimize an intein generating system in yeast instead of bacteria
|Marshall CJ,Grosskopf VA,Moehling TJ,Tillotson BJ,Wiepz GJ,Abbott NL,Raines RT,Shusta EV||ACS chemical biology (10:527)||2015|
|Human||Not Cited||Loss of Sh3gl2/endophilin A1 is a common event in urothelial carcinoma that promotes malignant behavior.||Majumdar S,Gong EM,Di Vizio D,Dreyfuss J,Degraff DJ,Hager MH,Park PJ,Bellmunt J,Matusik RJ,Rosenberg JE,Adam RM||Neoplasia (New York, N.Y.) (15:749)||2013|
||Sialyl Lewis X modification of the epidermal growth factor receptor regulates receptor function during airway epithelial wound repair.||Allahverdian S,Wang A,Singhera GK,Wong BW,Dorscheid DR||Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology (40:607)||2010|
A cell-based immunocytochemical assay for monitoring kinase signaling pathways and drug efficacy.
AHR5062 was used in ELISA to develop a near-infrared cytoblot assay and use it to study kinase signaling and the effects of kinase inhibitors
|Chen H,Kovar J,Sissons S,Cox K,Matter W,Chadwell F,Luan P,Vlahos CJ,Schutz-Geschwender A,Olive DM||Analytical biochemistry (338:136)||2005|