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 + 199.12) Mouse Monoclonal Antibody (MA513697) 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 membranous localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Rat, Non-human primate, Hamster, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Clone||528 + 199.12|
|Immunogen||Partially purified EGFR from A431 cells (clone 528) and extracellular domain of recombinant human EGFR protein (clone 199.12)|
|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)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||2 µg/ml|
* 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 2 publications below|
|ELISA (ELISA)||See 1 publications below|
|Immunoprecipitation (IP)||See 20 publications below|
|Immunocytochemistry (ICC)||See 3 publications below|
|Blocking Assay (BLOCK)||See 1 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
|Flow Cytometry (Flow)||See 1 publications below|
MA5-13697 targets Epidermal Growth Factor Receptor in IP applications and shows reactivity with Human samples.
The MA5-13697 immunogen is partially purified EGFR from A431 cells (clone 528) and extracellular domain of recombinant human EGFR protein (clone 199.12).
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 119-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. MA5-13697). 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.
Surfactant protein D suppresses lung cancer progression by downregulation of epidermal growth factor signaling.
MA5-13697 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|
Activation of Keap1/Nrf2 signaling pathway by nuclear epidermal growth factor receptor in cancer cells.
MA5-13697 was used in western blot to study the role of EGFR in regulating the Keap1/Nrf2 cascade
|Huo L,Li CW,Huang TH,Lam YC,Xia W,Tu C,Chang WC,Hsu JL,Lee DF,Nie L,Yamaguchi H,Wang Y,Lang J,Li LY,Chen CH,Mishra L,Hung MC||American journal of translational research (6:649)||2015|
High speed digital protein interaction analysis using microfluidic single molecule detection system.
MA5-13697 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.
MA5-13697 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.
MA5-13697 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|
Epidermal growth factor receptor signaling intensity determines intracellular protein interactions, ubiquitination, and internalization.
MA5-13697 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|
A sandwich type acridinium-linked immunosorbent assay (ALISA) detects soluble ErbB1 (sErbB1) in normal human sera.
MA5-13697 was used in western blot to develop an acridium-linked immunosorbent assay for measuring soluble ErbB1 in human serum
|Baron AT,Lafky JM,Connolly DC,Peoples J,O'Kane DJ,Suman VJ,Boardman CH,Podratz KC,Maihle NJ||Journal of immunological methods (219:23)||1998|
|Not Applicable||Not Cited||
PML represses lung cancer metastasis by suppressing the nuclear EGFR-mediated transcriptional activation of MMP2.
MA5-13697 was used in ChIP assay to study the suppression of nuclear EGFR-mediated transcriptional activation of MMP2 by PML that represses lung cancer metastasis
|Kuo HY,Huang YS,Tseng CH,Chen YC,Chang YW,Shih HM,Wu CW||Cell cycle (Georgetown, Tex.) (13:3132)||2014|
mMAPS: a flow-proteometric technique to analyze protein-protein interactions in individual signaling complexes.
MA5-13697 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|
A Miniaturized Ligand Binding Assay for EGFR.
MA5-13697 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.
MA5-13697 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|
Quantitative assays for the measurement of HER1-HER2 heterodimerization and phosphorylation in cell lines and breast tumors: applications for diagnostics and targeted drug mechanism of action.
MA5-13697 was used in immunoprecipitation to investigate the state of HER1-HER2 interaction and modification in cell lines and tumors
|DeFazio-Eli L,Strommen K,Dao-Pick T,Parry G,Goodman L,Winslow J||Breast cancer research : BCR (13:null)||2011|
IL-24 is expressed during wound repair and inhibits TGFalpha-induced migration and proliferation of keratinocytes.
MA5-13697 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.
MA5-13697 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|
Ubc4/5 and c-Cbl continue to ubiquitinate EGF receptor after internalization to facilitate polyubiquitination and degradation.
MA5-13697 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|
Ligand release-independent transactivation of epidermal growth factor receptor by transforming growth factor-beta involves multiple signaling pathways.
MA5-13697 was used in immunoprecipitation to examine the involvement of epidermal growth factor receptor in transforming growth factor-beta signaling pathways
|Joo CK,Kim HS,Park JY,Seomun Y,Son MJ,Kim JT||Oncogene (27:614)||2008|
Impact of common epidermal growth factor receptor and HER2 variants on receptor activity and inhibition by lapatinib.
MA5-13697 was used in immunoprecipitation to investigate the effect of lapatinib on cell proliferation regulated by EGFR/HER2
|Gilmer TM,Cable L,Alligood K,Rusnak D,Spehar G,Gallagher KT,Woldu E,Carter HL,Truesdale AT,Shewchuk L,Wood ER||Cancer research (68:571)||2008|
|Non-human primate||Not Cited||
Sensitivity of epidermal growth factor receptor and ErbB2 exon 20 insertion mutants to Hsp90 inhibition.
MA5-13697 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.
MA5-13697 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.
MA5-13697 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.
MA5-13697 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.
MA5-13697 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|
A unique structure for epidermal growth factor receptor bound to GW572016 (Lapatinib): relationships among protein conformation, inhibitor off-rate, and receptor activity in tumor cells.
MA5-13697 was used in immunoprecipitation to conduct the structural analysis of the binding of epidermal growth factor receptor to GW572016
|Wood ER,Truesdale AT,McDonald OB,Yuan D,Hassell A,Dickerson SH,Ellis B,Pennisi C,Horne E,Lackey K,Alligood KJ,Rusnak DW,Gilmer TM,Shewchuk L||Cancer research (64:6652)||2004|
Autocrine extracellular signal-regulated kinase (ERK) activation in normal human keratinocytes: metalloproteinase-mediated release of amphiregulin triggers signaling from ErbB1 to ERK.
MA5-13697 was used in immunoprecipitation to investigate the mechanism of ERK autocrine activation in normal human keratinocytes
|Kansra S,Stoll SW,Johnson JL,Elder JT||Molecular biology of the cell (15:4299)||2004|
Geldanamycin-associated inhibition of intracellular trafficking is attributed to a co-purified activity.
MA5-13697 was used in immunoprecipitation to study a co-purifying contaminant of geldanamycin preparations as the component inhibiting intracellular trafficking
|Barzilay E,Ben-Califa N,Supino-Rosin L,Kashman Y,Hirschberg K,Elazar Z,Neumann D||The Journal of biological chemistry (279:6847)||2004|
1,25-Dihydroxyvitamin D down-regulates cell membrane growth- and nuclear growth-promoting signals by the epidermal growth factor receptor.
MA5-13697 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|
Epidermal growth factor receptor signaling and the invasive phenotype of ovarian carcinoma cells.
MA5-13697 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.
MA5-13697 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|
Intracellular retention and degradation of the epidermal growth factor receptor, two distinct processes mediated by benzoquinone ansamycins.
MA5-13697 was used in immunoprecipitation to study the effect of the benzoquinone ansamycins on the intracellular retention and degradation of EGFR
|Supino-Rosin L,Yoshimura A,Yarden Y,Elazar Z,Neumann D||The Journal of biological chemistry (275:21850)||2000|
Epidermal growth factor receptor vIII enhances tumorigenicity in human breast cancer.
MA5-13697 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|
Monitoring the size and lateral dynamics of ErbB1 enriched membrane domains through live cell plasmon coupling microscopy.
MA5-13697 was used in immunocytochemistry to study the dynamics of ErbB1 enriched membrane domains using live cell plasmon coupling microscopy
|Rong G,Reinhard BM||PloS one (7:null)||2012|
Pgrmc1 (progesterone receptor membrane component 1) associates with epidermal growth factor receptor and regulates erlotinib sensitivity.
MA5-13697 was used in immunocytochemistry to investigate the association between progesterone receptor membrane component 1 and epidermal growth factor receptor and its effect on erlotinib sensitivity
|Ahmed IS,Rohe HJ,Twist KE,Craven RJ||The Journal of biological chemistry (285:24775)||2010|
The mucin Muc4 potentiates neuregulin signaling by increasing the cell-surface populations of ErbB2 and ErbB3.
MA5-13697 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|
Receptor tyrosine kinase inhibitor profiling using bead-based multiplex sandwich immunoassays.
MA5-13697 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|
Potential localization of putative stem/progenitor cells in human bulbar conjunctival epithelium.
MA5-13697 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|
Functional effects of glycosylation at Asn-579 of the epidermal growth factor receptor.
MA5-13697 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|