Immunofluorescence analysis of EGFR was done on 90% confluent log phase A431 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 EGFR (528) Mouse Monoclonal Antibody (MA512875) at 2ug/mL 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) at 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 (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Dog, Human|
|Published species reactivity||Rat, Non-human primate, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified EGFR from A431 cells|
|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)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:400|
|Immunoprecipitation (IP)||Assay Dependent|
* 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.
|Western Blot (WB)||See 7 publications below|
|ChIP assay (ChIP)||See 1 publications below|
|Blocking Assay (BLOCK)||See 2 publications below|
|Immunocytochemistry (ICC)||See 3 publications below|
|Flow Cytometry (Flow)||See 7 publications below|
|ELISA (ELISA)||See 1 publications below|
|Immunoprecipitation (IP)||See 16 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
MA5-12875 targets Epidermal Growth Factor Receptor in FACS, IHC and IP applications and shows reactivity with Canine and Human samples.
The MA5-12875 immunogen is purified EGFR from A431 cells.
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. Multiple alternatively spliced transcript variants that encode different protein isoforms have been found for this gene.
IP-MS enrichment of EGFR (LFQ intensity): EGFR was enriched 47-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. MA5-12875). 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.
Targeting a Cancer-Specific Epitope of the Epidermal Growth Factor Receptor in Triple-Negative Breast Cancer.
MA5-12875 was used in flow cytometry and western blot to characterize the epidermal growth factor receptor in triple-negative breast cancer for a cancer-specific epitope target
|Simon N,Antignani A,Sarnovsky R,Hewitt SM,FitzGerald D||Journal of the National Cancer Institute (108:null)||2016|
Surfactant protein D suppresses lung cancer progression by downregulation of epidermal growth factor signaling.
MA5-12875 was used in immunoprecipitation and western blot to study the role of impaired EGF binding to EGFR in the mechanism by which surfactant protein D inhibits the progression of lung cancer
|Hasegawa Y,Takahashi M,Ariki S,Asakawa D,Tajiri M,Wada Y,Yamaguchi Y,Nishitani C,Takamiya R,Saito A,Uehara Y,Hashimoto J,Kurimura Y,Takahashi H,Kuroki Y||Oncogene (34:838)||2015|
High speed digital protein interaction analysis using microfluidic single molecule detection system.
MA5-12875 was used in western blot to study the use of a microfluidic single molecule detection system for the rapid analysis of protein interactions
|Chou CK,Jing N,Yamaguchi H,Tsou PH,Lee HH,Chen CT,Wang YN,Hong S,Su C,Kameoka J,Hung MC||Lab on a chip (10:1793)||2010|
EGFRvIII-induced estrogen-independence, tamoxifen-resistance phenotype correlates with PgR expression and modulation of apoptotic molecules in breast cancer.
MA5-12875 was used in western blot to study the ability of EGFRvIII to promote estrogen-independent growth in breast cancer cell lines
|Zhang Y,Su H,Rahimi M,Tochihara R,Tang C||International journal of cancer (125:2021)||2009|
EGFR and ADAMs cooperate to regulate shedding and endocytic trafficking of the desmosomal cadherin desmoglein 2.
MA5-12875 was used in western blot to examine the role of EGFR and ADAMs in the endocytic trafficking of the desmosomal cadherin desmoglein 2
|Klessner JL,Desai BV,Amargo EV,Getsios S,Green KJ||Molecular biology of the cell (20:328)||2009|
LRIG1 negatively regulates the oncogenic EGF receptor mutant EGFRvIII.
MA5-12875 was used in western blot to examine the role of LRIG1 in the degradation of oncogenic EGF receptor mutant EGFRvIII
|Stutz MA,Shattuck DL,Laederich MB,Carraway KL,Sweeney C||Oncogene (27:5741)||2008|
Epidermal growth factor receptor signaling intensity determines intracellular protein interactions, ubiquitination, and internalization.
MA5-12875 was used in western blot to study the role of EGFR signaling intensity in determining intracellular protein interactions, ubiquitination, and internalization
|Schmidt MH,Furnari FB,Cavenee WK,Bögler O||Proceedings of the National Academy of Sciences of the United States of America (100:6505)||2003|
mMAPS: a flow-proteometric technique to analyze protein-protein interactions in individual signaling complexes.
MA5-12875 was used in ChIP assay to develop a novel flow-proteomic methodology for the detection of individual signaling complex protein-protein interactions in crude cell/tissue lysates
|Chou CK,Lee HH,Tsou PH,Chen CT,Hsu JM,Yamaguchi H,Wang YN,Lee HJ,Hsu JL,Lee JF,Kameoka J,Hung MC||Science signaling (7:null)||2014|
Propranolol restricts the mobility of single EGF-receptors on the cell surface before their internalization.
MA5-12875 was used in blocking or activating experiment to study the ability of propanolol to restrict cell surface EGFR mobility prior to receptor internalization
|Otero C,Linke M,Sanchez P,González A,Schaap IA||PloS one (8:null)||2013|
Receptor tyrosine kinase inhibitor profiling using bead-based multiplex sandwich immunoassays.
MA5-12875 was used in blocking or activating experiment to develop a bead-based sandwich immunoassay for the profiling of receptor tyrosine kinase inhibitors
|Pötz O,Schneiderhan-Marra N,Henzler T,Herget T,Joos TO||Methods in molecular biology (Clifton, N.J.) (795:191)||2011|
Internalization mechanisms of the epidermal growth factor receptor after activation with different ligands.
MA5-12875 was used in immunocytochemistry to study internalization of the EGFR following activation by various ligands and the different mechanisms involved
|Henriksen L,Grandal MV,Knudsen SL,van Deurs B,Grøvdal LM||PloS one (8:null)||2013|
The mucin Muc4 potentiates neuregulin signaling by increasing the cell-surface populations of ErbB2 and ErbB3.
MA5-12875 was used in immunocytochemistry to study the role of ErbB2 and ErbB3 in the mechanism by which Muc4 potentiates neuregulin signaling
|Funes M,Miller JK,Lai C,Carraway KL,Sweeney C||The Journal of biological chemistry (281:19310)||2006|
Suppression of EGFRvIII-mediated proliferation and tumorigenesis of breast cancer cells by ribozyme.
MA5-12875 was used in immunocytochemistry to evaluate the anti-tumor effect of tumor specific ribozyme targeted to EGFRvIII
|Luo X,Gong X,Tang CK||International journal of cancer (104:716)||2003|
Intracytoplasmic epidermal growth factor receptor shows poor response to the cetuximab antitumor effect in irradiated non-small cell lung cancer cell lines.
MA5-12875 was used in flow cytometry to study the anti-tumor activity of a blocking antibody against cytoplasmic EGFR in a non-small cell lung cancer cell line
|Ryu SH,Lee SW,Yang YJ,Song SY,Kim JH,Choi EK,Ahn SD||Lung cancer (Amsterdam, Netherlands) (77:482)||2012|
The oncoprotein ErbB3 is endocytosed in the absence of added ligand in a clathrin-dependent manner.
MA5-12875 was used in flow cytometry to study the clathrin-dependent endocytosis of the ErbB3 oncoprotein
|Sak MM,Breen K,Rønning SB,Pedersen NM,Bertelsen V,Stang E,Madshus IH||Carcinogenesis (33:1031)||2012|
Molecular basis for sensitivity and acquired resistance to gefitinib in HER2-overexpressing human gastric cancer cell lines derived from liver metastasis.
MA5-12875 was used in flow cytometry to study the possibility of molecular therapy targeting HER2 overexpression in gastric cancer liver metastasis
|Yokoyama H,Ikehara Y,Kodera Y,Ikehara S,Yatabe Y,Mochizuki Y,Koike M,Fujiwara M,Nakao A,Tatematsu M,Nakanishi H||British journal of cancer (95:1504)||2006|
Directed evolution of the epidermal growth factor receptor extracellular domain for expression in yeast.
MA5-12875 was used in flow cytometry to express functional and properly folded EGFR extracellular domain in yeast
|Kim YS,Bhandari R,Cochran JR,Kuriyan J,Wittrup KD||Proteins (62:1026)||2006|
Functional effects of glycosylation at Asn-579 of the epidermal growth factor receptor.
MA5-12875 was used in flow cytometry to study the functional effects of N-glycosylation of Asn-579 of EGFR
|Whitson KB,Whitson SR,Red-Brewer ML,McCoy AJ,Vitali AA,Walker F,Johns TG,Beth AH,Staros JV||Biochemistry (44:14920)||2005|
Domain-level antibody epitope mapping through yeast surface display of epidermal growth factor receptor fragments.
MA5-12875 was used in flow cytometry to study the use of yeast surface display for identifying stable folded protein domains and characterizing antibody binding epitopes
|Cochran JR,Kim YS,Olsen MJ,Bhandari R,Wittrup KD||Journal of immunological methods (287:147)||2004|
Ribozyme-mediated down-regulation of ErbB-4 in estrogen receptor-positive breast cancer cells inhibits proliferation both in vitro and in vivo.
MA5-12875 was used in flow cytometry to study the role of ErbB-4 in the proliferation of estrogen receptor-positive breast cancer cells
|Tang CK,Concepcion XZ,Milan M,Gong X,Montgomery E,Lippman ME||Cancer research (59:5315)||1999|
A Miniaturized Ligand Binding Assay for EGFR.
MA5-12875 was used in ELISA to develop a miniaturized ligand binding assay for EGFR
|Schwenk JM,Poetz O,Zeillinger R,Joos TO||International journal of proteomics (2012:null)||2012|
Superior antitumoral activity of dimerized targeted single-chain TRAIL fusion proteins under retention of tumor selectivity.
MA5-12875 was used in immunoprecipitation to develop targeted dimerized single-chain TRAIL fusion proteins displaying markedly increased anti-tumor activity
|Siegemund M,Pollak N,Seifert O,Wahl K,Hanak K,Vogel A,Nussler AK,Göttsch D,Münkel S,Bantel H,Kontermann RE,Pfizenmaier K||Cell death and disease (3:null)||2012|
IL-24 is expressed during wound repair and inhibits TGFalpha-induced migration and proliferation of keratinocytes.
MA5-12875 was used in immunoprecipitation to study the role of IL-24 in wound repair and its inhibitory effect on TGFalpha-induced migration and proliferation of keratinocytes
|Poindexter NJ,Williams RR,Powis G,Jen E,Caudle AS,Chada S,Grimm EA||Experimental dermatology (19:714)||2010|
Intracellular MUC1 peptides inhibit cancer progression.
MA5-12875 was used in immunoprecipitation to study the anti-tumor activity of cell-permeant peptides designed to block MUC1 interactions with beta-catenin and EGFR
|Bitler BG,Menzl I,Huerta CL,Sands B,Knowlton W,Chang A,Schroeder JA||Clinical cancer research : an official journal of the American Association for Cancer Research (15:100)||2009|
Freud-1/Aki1, a novel PDK1-interacting protein, functions as a scaffold to activate the PDK1/Akt pathway in epidermal growth factor signaling.
MA5-12875 was used in immunoprecipitation to investigate the role of Freud-1/Aki1 in the PDK1/Akt pathway
|Nakamura A,Naito M,Tsuruo T,Fujita N||Molecular and cellular biology (28:5996)||2008|
Ubc4/5 and c-Cbl continue to ubiquitinate EGF receptor after internalization to facilitate polyubiquitination and degradation.
MA5-12875 was used in immunoprecipitation to study the role of Ubc4/5 and c-Cbl in the polyubiquitination and degradation of internalized EGFR
|Umebayashi K,Stenmark H,Yoshimori T||Molecular biology of the cell (19:3454)||2008|
|Non-human primate||Not Cited||
Sensitivity of epidermal growth factor receptor and ErbB2 exon 20 insertion mutants to Hsp90 inhibition.
MA5-12875 was used in immunoprecipitation to study the sensitivity of EGFR and ErbB2 exon 20 insertion mutants to Hsp90 inhibition
|Xu W,Soga S,Beebe K,Lee MJ,Kim YS,Trepel J,Neckers L||British journal of cancer (97:741)||2007|
Acquired resistance to erlotinib in A-431 epidermoid cancer cells requires down-regulation of MMAC1/PTEN and up-regulation of phosphorylated Akt.
MA5-12875 was used in immunoprecipitation to study the mechanism of acquired resistance to erlotinib in epidermoid cancer cells
|Yamasaki F,Johansen MJ,Zhang D,Krishnamurthy S,Felix E,Bartholomeusz C,Aguilar RJ,Kurisu K,Mills GB,Hortobagyi GN,Ueno NT||Cancer research (67:5779)||2007|
Intersectin regulates epidermal growth factor receptor endocytosis, ubiquitylation, and signaling.
MA5-12875 was used in immunoprecipitation to study the mechanism by which the endocytic scaffold protein intersectin regulates EGFR degradation and signaling
|Martin NP,Mohney RP,Dunn S,Das M,Scappini E,O'Bryan JP||Molecular pharmacology (70:1643)||2006|
Diacylglycerol kinase delta regulates protein kinase C and epidermal growth factor receptor signaling.
MA5-12875 was used in immunoprecipitation to study the role of PKC in the mechanism by which diacylglycerol kinase-delta regulates EGFR signaling
|Crotty T,Cai J,Sakane F,Taketomi A,Prescott SM,Topham MK||Proceedings of the National Academy of Sciences of the United States of America (103:15485)||2006|
Nuclear-cytoplasmic transport of EGFR involves receptor endocytosis, importin beta1 and CRM1.
MA5-12875 was used in immunoprecipitation to study the mechanism of EGFR nuclear-cytoplasmic transport
|Lo HW,Ali-Seyed M,Wu Y,Bartholomeusz G,Hsu SC,Hung MC||Journal of cellular biochemistry (98:1570)||2006|
1,25-Dihydroxyvitamin D down-regulates cell membrane growth- and nuclear growth-promoting signals by the epidermal growth factor receptor.
MA5-12875 was used in immunoprecipitation to study the down regulatory effects of 1,25-dihydroxyvitamin D on EGFR membrane trafficing and growth signaling
|Cordero JB,Cozzolino M,Lu Y,Vidal M,Slatopolsky E,Stahl PD,Barbieri MA,Dusso A||The Journal of biological chemistry (277:38965)||2002|
Metalloproteinases stimulate ErbB-dependent ERK signaling in human skin organ culture.
MA5-12875 was used in immunoprecipitation to study the role of MMPs in stimulating ErbB-dependent ERK signaling in human skin organ culture
|Stoll SW,Kansra S,Elder JT||The Journal of biological chemistry (277:26839)||2002|
Epidermal growth factor receptor signaling and the invasive phenotype of ovarian carcinoma cells.
MA5-12875 was used in immunoprecipitation to study the role of EGFR signaling in the invasiveness of ovarian carcinoma cells
|Bergmann-Leitner ES,Bennett TA,Hacker NF,Stromberg K,Stetler-Stevenson WG||Journal of the National Cancer Institute (93:1375)||2001|
Tyrosine-phosphorylated plakoglobin is associated with desmogleins but not desmoplakin after epidermal growth factor receptor activation.
MA5-12875 was used in immunoprecipitation to study the effects of EGFR activation on the association of tyrosine phosphorylated plakoglobin with desmogleins and desmoplakin
|Gaudry CA,Palka HL,Dusek RL,Huen AC,Khandekar MJ,Hudson LG,Green KJ||The Journal of biological chemistry (276:24871)||2001|
Epidermal growth factor receptor vIII enhances tumorigenicity in human breast cancer.
MA5-12875 was used in immunoprecipitation to study the role of the constitutively active EGFR vIII variant in the proliferation and tumorigenicity of human breast cancer
|Tang CK,Gong XQ,Moscatello DK,Wong AJ,Lippman ME||Cancer research (60:3081)||2000|
Mitosis-specific negative regulation of epidermal growth factor receptor, triggered by a decrease in ligand binding and dimerization, can be overcome by overexpression of receptor.
MA5-12875 was used in immunoprecipitation and western blot to study the mechanism of the mitosis-specific negative regulation of EGFR and the effect receptor overexpression
|Kiyokawa N,Lee EK,Karunagaran D,Lin SY,Hung MC||The Journal of biological chemistry (272:18656)||1997|
Potential localization of putative stem/progenitor cells in human bulbar conjunctival epithelium.
MA5-12875 was used in immunohistochemistry to study the expression pattern of stem cells in the bulbar conjunctival epithelium
|Qi H,Zheng X,Yuan X,Pflugfelder SC,Li DQ||Journal of cellular physiology (225:180)||2010|