Overview

  • Product name
    Anti-EGFR antibody [ICR10]
    See all EGFR primary antibodies
  • Description
    Rat monoclonal [ICR10] to EGFR
  • Host species
    Rat
  • Tested applications
    Suitable for: Inhibition Assay, Flow Cyt, ELISA, ICC/IF, IP, IHC-Fr, ICCmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

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

  • Epitope
    ICR10 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.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Function
      Receptor 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 specificity
      Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
    • Involvement in disease
      Lung cancer
      Inflammatory skin and bowel disease, neonatal, 2
    • Sequence similarities
      Belongs 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. Dephosphorylated by PTPN1 and PTPN2.
      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 occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
      Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.
    • Cellular localization
      Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF).
    • Information by UniProt
    • Database links
    • Alternative names
      • Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
      • Cell growth inhibiting protein 40 antibody
      • Cell proliferation inducing protein 61 antibody
      • EGF R 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
      • erb-b2 receptor tyrosine kinase 1 antibody
      • ERBB antibody
      • ERBB1 antibody
      • Errp antibody
      • HER1 antibody
      • mENA antibody
      • NISBD2 antibody
      • Oncogen ERBB antibody
      • PIG61 antibody
      • Proto-oncogene c-ErbB-1 antibody
      • Receptor tyrosine protein kinase ErbB 1 antibody
      • Receptor tyrosine-protein kinase ErbB-1 antibody
      • SA7 antibody
      • Species antigen 7 antibody
      • Urogastrone antibody
      • v-erb-b Avian erythroblastic leukemia viral oncogen homolog antibody
      • wa2 antibody
      • Wa5 antibody
      see all

    Images

    • 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

    • 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.

    References

    This product has been referenced in:
    • Gerlach JP  et al. Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells. Sci Rep 9:1469 (2019). Read more (PubMed: 30728416) »
    • Ortega FG  et al. EGFR detection in extracellular vesicles of breast cancer patients through immunosensor based on silica-chitosan nanoplatform. Talanta 194:243-252 (2019). Read more (PubMed: 30609526) »
    See all 17 Publications for this product

    Customer reviews and Q&As

    Filter by Application

    Filter by Species

    Filter by Ratings

    1-5 of 5 Abreviews

    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

    Dr. Jennifer Dembinski

    Verified customer

    Submitted May 14 2009

    This product is known to not work in this application or species.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A431 cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Specification
    A431 cells
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Aug 16 2017

    Application
    Flow Cytometry
    Sample
    Human Cell (HCC827 cells)
    Permeabilization
    No
    Gating Strategy
    Live cells (PI negative)
    Specification
    HCC827 cells
    Preparation
    Cell harvesting/tissue preparation method: Trypsinized
    Sample buffer: RPMI1640 with 10% FBS and Penicillin-Streptomycin
    Fixation
    No

    Abcam user community

    Verified customer

    Submitted Jul 14 2017

    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

    Abcam user community

    Verified customer

    Submitted Feb 24 2012

    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

    Ms. Lakshmi Ganesan

    Verified customer

    Submitted Nov 20 2009

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