Overview
- Product nameAnti-EGFR antibody [2E9]See all EGFR primary antibodies ...
- DescriptionMouse monoclonal [2E9] to EGFR
- SpecificityThis antibody reacts with a protein determinant of the extra cellular domain of the human EGFR. Shows binding competition with EGF and reacts in immunoprecipitation with the functional EFR protein-tyrosine kinase complex.
- Tested applicationsImmunomicroscopy, IP, ICC/IF more details
- Species reactivityReacts with: Human
Does not react with
Mouse - Positive controlMetastasis lung carcinoma Adenocarcinoma rectum Epithelium tabular L carcinoma
- General notesSufficient for 100-200 tests
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
- PurityTissue culture supernatant
- Clonality Monoclonal
- Clone number2E9
- Myelomaunknown
- IsotypeIgG1
- Light chain typeunknown
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Research Areas
Applications
Our Abpromise guarantee covers the use of ab8465 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| Immunomicroscopy | |
| IP | |
| ICC/IF |
IM: Use at an assay dependent dilution.
IP: Use at an assay dependent dilution.
Is unsuitable for IHC-P or IHC-Fr.
The antibody has no agonistic properties, but competes efficiently with EGF for binding to the receptor.
Optimal dilutions/concentrations should be determined by the end user.
Target
- FunctionReceptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
Isoform 2 may act as an antagonist of EGF action. - Tissue specificityUbiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
- Involvement in diseaseDefects in EGFR are associated with lung cancer (LNCR) [MIM:211980]. LNCR is a common malignancy affecting tissues of the lung. The most common form of lung cancer is non-small cell lung cancer (NSCLC) that can be divided into 3 major histologic subtypes: squamous cell carcinoma, adenocarcinoma, and large cell lung cancer. NSCLC is often diagnosed at an advanced stage and has a poor prognosis.
- Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. - Post-translational
modificationsPhosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3.
Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occur. Deubiquitinated by OTUD7B, preventing degradation.
Methylated. Methylation at Arg-1199 by PRMT5 positively stimulates phosphorylation at Tyr-1197. - Cellular localizationSecreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand.
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Database links
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
Target information above from: UniProt accession
P00533
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
.
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Alternative names
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibodyAvian erythroblastic leukemia viral (verbb) oncogene homolog antibodyCell growth inhibiting protein 40 antibody
- Cell proliferation inducing protein 61 antibodyEGF R antibodyEGFR antibodyEGFR antibodyEGFR_HUMAN antibodyEpidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) antibodyEpidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) antibodyEpidermal growth factor receptor antibodyerbb 1 antibodyErbb antibodyErbb1 antibodyERBB1 antibodyErrp antibodyHER1 antibodymENA antibodyOncogene ERBB antibodyPIG61 antibodyProto-oncogene c-ErbB-1 antibodyReceptor tyrosine protein kinase ErbB 1 antibodyReceptor tyrosine protein kinase ErbB1 antibodyReceptor tyrosine-protein kinase ErbB-1 antibodyUrogastrone antibodywa2 antibodyWa5 antibody
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Anti-EGFR antibody [2E9] images
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ICC/IF image of ab8465 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8465, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
References for Anti-EGFR antibody [2E9] (ab8465)
This product has been referenced in:
- Defize LH et al. Signal transduction by epidermal growth factor occurs through the subclass of high affinity receptors. J Cell Biol 109:2495-507 (1989). Read more (PubMed: 2553748) »
![Immunocytochemistry/ Immunofluorescence - EGFR antibody [2E9] (ab8465) Immunocytochemistry/ Immunofluorescence - EGFR antibody [2E9] (ab8465)](http://a.abcam.com/ps/datasheet/images/8/ab8465/EGFR-Primary-antibodies-ab8465-2.jpg)