Overview

  • Product nameAnti-EGFR [2E9] antibodySee all EGFR primary antibodies ...
  • Description
    Mouse 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. The antibody has no agonistic properties, but competes efficiently with EGF for binding to the receptor
  • Tested applicationsImmunomicroscopy, IP, ICC/IF, Flow Cyt more details
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse
  • Positive control
    • Metastasis lung carcinoma Adenocarcinoma rectum Epithelium tabular L carcinoma
  • General notesSufficient for 100-200 tests

Properties

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 Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF 1/1000.
Flow Cyt Use 0.1µg for 106 cells.
  • Application notesIs unsuitable for IHC-Fr or IHC-P.
  • 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
      modifications
      Phosphorylation 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.
    • Target information above from: UniProt accession P00533 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links
    • Alternative names
      • Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
      • Avian erythroblastic leukemia viral (verbb) oncogene homolog antibody
      • Cell growth inhibiting protein 40 antibody
      • Cell proliferation inducing protein 61 antibody
      • EGF R antibody
      • EGFR antibody
      • EGFR antibody
      • EGFR_HUMAN antibody
      • Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) antibody
      • Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) antibody
      • Epidermal growth factor receptor antibody
      • erbb 1 antibody
      • Erbb antibody
      • Erbb1 antibody
      • ERBB1 antibody
      • Errp antibody
      • HER1 antibody
      • mENA antibody
      • Oncogene ERBB antibody
      • PIG61 antibody
      • Proto-oncogene c-ErbB-1 antibody
      • Receptor tyrosine protein kinase ErbB 1 antibody
      • Receptor tyrosine protein kinase ErbB1 antibody
      • Receptor tyrosine-protein kinase ErbB-1 antibody
      • Urogastrone antibody
      • wa2 antibody
      • Wa5 antibody
      see all

    Anti-EGFR [2E9] antibody images

    • Overlay histogram showing A431 cells stained with ab8465 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8465, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • 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.

    References for Anti-EGFR [2E9] antibody (ab8465)

    This product has been referenced in:
    • Zanetti-Domingues LC  et al. Hydrophobic fluorescent probes introduce artifacts into single molecule tracking experiments due to non-specific binding. PLoS One 8:e74200 (2013). ICC/IF . Read more (PubMed: 24066121) »
    • 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) »

    See all 2 Publications for this product

    Product Wall

    Thank you for your email.

    The product ab8465 is specific to extra cellular domain of the receptor.

    Thanks!

    Thank you for your enquiry. I have been in touch with the source of this antibody and I can tell you that no pre-treatment was used by the developers of this clone to test the antibody on IHC-paraffin sections. I hope this information helps, pl...

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    Thank you for taking the time to contact me with the problems that you have experienced using this product in IHC-P. Looking through the information that you have forwarded there are a few suggestions that I would make that may improve the results th...

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    Unfortunately, we are not specialists in this area and we do not want to recommend a product that is unsuitable for your needs. We suggest that you refer to the latest literature in this field.

    We have no further information on the cross reactivity of this antibody with other isoforms of the EGF-Receptor.

    I'm afraid that I cannot be of much help. All the information we have for this antibody is on the datasheet where you can see that it has no agonistic properties, but competes efficiently with EGF for binding to the receptor. We know that the antibody ...

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    No, the epitope recognised by this antibody has not been mapped further

    This antibody works in a lot of applications , but it has not yet been tested in WB.

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"