Overview

  • Product nameAnti-EGFR antibody [ICR10]See all EGFR primary antibodies ...
  • Description
    Rat monoclonal [ICR10] to EGFR
  • SpecificityICR10 recognises only wild-type Human epidermal growth factor receptor (EGF-R). It does not react with erb-B2.
  • Tested applicationsInhibition Assay, Flow Cyt, ELISA, IP, IHC-Fr, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Extracellular domain of Human EGF-receptor from head and neck carcinoma

  • EpitopeICR10 binds to epitope B, and has an affinity of 6.7x10-9 M.
  • Positive control
    • FACS: A431 cells IHC: Breast Carcinoma tissue WB: HN5 cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab231 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Inhibition Assay Use at an assay dependent concentration. See ELISA Abreview.
Flow Cyt 1/50 - 1/100.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
  • Application notesIs unsuitable for WB.
  • 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.
    • 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 antibody [ICR10] images

    • Staining of A431 cells with ab231 visualised with F(ab')2 rabbit anti-rat FITC conjugated secondary.
    • ICC/IF image of ab231 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab231, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    • ab231 used undiluted in inhibition assay against human recombinant protein EGFR1:Fc to inhibit binding to EGF:Biotin. The primary incubated with sample for 1 hour. The antibody inhibits the binding of biotinylated EGF to coated EGFR:Fc with an IC50 ~ 5 nM.

    • ab231 staining EGFR in human tumor cell line xenograft in mouse by Immunohistochemistry (Frozen sections).
      Tissue was fixed in acetone. Samples incubated with ab231 at a 1/100 dilution for 1 hour at 23°C. The secondary used was an undiluted HRP conjugated goat anti-rat polyclonal.

      See Abreview

    References for Anti-EGFR antibody [ICR10] (ab231)

    This product has been referenced in:
    • Wang J & Yu RK Interaction of ganglioside GD3 with an EGF receptor sustains the self-renewal ability of mouse neural stem cells in vitro. Proc Natl Acad Sci U S A 110:19137-42 (2013). Flow Cyt, ICC/IF ; Mouse . Read more (PubMed: 24198336) »
    • Geißler C  et al. The challenge of tumor heterogeneity--different phenotypes of cancer stem cells in a head and neck squamous cell carcinoma xenograft mouse model. In Vivo 26:593-8 (2012). Read more (PubMed: 22773573) »

    See all 7 Publications for this product

    Product Wall

    Thank you for your inquiry regarding ab231.
    We do not measure the isoelectric point of our antibodies. Most monoclonal antibodies will have an isoelectric point around 6.1, but this is not something that we test.
    Please let me know if you hav...

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    Thank you very much for your reply.

    I've looked at the package'stracking information, and it arrived on September 20 at 9:34 EST. You can track the package on FedEx.com with tracking number: ***. It was signed for by ***- does that help you t...

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    Thank you very much for your reply.

    It does sound like the secondary is reacting with the IgG's in the mouse serum. I'm sending a free of charge vial of ab6115 on the order *** which should arrive tomorrow.

    Please keep me updated ab...

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    Thank you very much for contacting us with your question.

    It is possible that the secondary antibody is causing some background signal, as ab7100 has been pre-adsorbed only against human serum proteins and not mouse serum proteins. Have you d...

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    Thank you for contacting us.

    A common fluorophore used is FITC (green), and we carry ab7100 a rabbit polyclonal secondary antibody to rat iGG - H&L (FITC), pre-adsorbed as ab7100. This has been tested in FC.

    http://www.abcam.co...

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    Thank you for your enquiry and your interest in our products.

    I can confirm that neither of these antibodies have been characterized for inhibition studies. ab24284 is suitable for IHC-P only; whilst ab24293 has been tested for Flow cytometry...

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Sample Human Tissue sections (tumor cell line xenograft in mouse)
    Specification tumor cell line xenograft in mouse
    Fixative Acetone
    Permeabilization No
    Username

    Abcam user community

    Verified customer

    Submitted Feb 24 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application ELISA
    Sample Human Recombinant protein (EGFR1:Fc)
    Specification EGFR1:Fc
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Type Other
    Username

    Ms. Lakshmi Ganesan

    Verified customer

    Submitted Nov 20 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Sample Mouse Tissue sections (CB17/SCID BxPC-3 xenograft)
    Specification CB17/SCID BxPC-3 xenograft
    Fixative Paraformaldehyde
    Permeabilization No
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Dr. Jennifer Dembinski

    Verified customer

    Submitted May 14 2009

    Thank you for your enquiry. I am not sure I understand your question. Ab231 is a mouse monoclonal purified IgG prepared by ion exchange chromotography from ascites. Ab3103 is protein A purified, but I do not have the information on whether this wa...

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