Overview

  • Product name
    Anti-EGFR antibody [EGFR1]
    See all EGFR primary antibodies
  • Description
    Mouse monoclonal [EGFR1] to EGFR
  • Host species
    Mouse
  • Specificity

    Recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR. This antibody does not react with erbB-2, erbB-3 and erbB-4.

  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, IP, Flow Cyt, IHC - Wholemountmore details
    Unsuitable for: ELISA or WB
  • Species reactivity
    Reacts with: Mouse, Horse, Human
  • Immunogen

    Tissue, cells or virus corresponding to Human EGFR (extracellular). Human epidermoid carcinoma line A431; epitope mapped between aa 6-273 of human EGFR.

  • Epitope
    Extracellular.
  • Positive control
    • IHC-Fr: Frozen normal human placenta ICC/IF: A431 cells Flow Cyt: A 431 cells
  • General notes

    Recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR.

    This antibody clone is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab30 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
IHC-Fr Use a concentration of 10 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
Flow Cyt Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

IHC - Wholemount Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ELISA or 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

    • Overlay histograms showing left positive A431 cells and right negative Jurkat cells stained with ab30 (red line). The cells were incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab30, 1x106 cells in 100ul at 1ug/ml) for 30 min on ice. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) preabsorbed (ab150117) at 1/2000 dilution for 30 min on ice.

      Isotype control antibody (black line) was mouse IgG2bκ (ab170192) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    • ab30 staining EGFR (colored green) in A431 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab30 at 5μg/ml overnight at 4ºC. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • IHC image of EGFR staining in a section of frozen normal human placenta, fixed in 10% paraformaldehyde (10 min). Staining was performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab30, 10ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References

    This product has been referenced in:
    • Fisher SA  et al. Photo-immobilized EGF chemical gradients differentially impact breast cancer cell invasion and drug response in defined 3D hydrogels. Biomaterials 178:751-766 (2018). Flow Cyt . Read more (PubMed: 29452913) »
    • Minutti CM  et al. Epidermal Growth Factor Receptor Expression Licenses Type-2 Helper T Cells to Function in a T Cell Receptor-Independent Fashion. Immunity 47:710-722.e6 (2017). Read more (PubMed: 29045902) »
    See all 16 Publications for this product

    Customer reviews and Q&As

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    1-4 of 4 Abreviews

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human tissue from different sites (TMA))
    Antigen retrieval step
    Other - Buffer/Enzyme Used: Citrate pH6, Tris-EDTA pH 9 and Chymotrypsin
    Permeabilization
    No
    Specification
    Human tissue from different sites (TMA)
    Blocking step
    (agent) for 10 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Formaldehyde

    Mr. Malcolm Lim

    Verified customer

    Submitted Jul 14 2017

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Breast cancer cell lines)
    Permeabilization
    Yes - 0.5% tritonx/PBS
    Specification
    Breast cancer cell lines
    Blocking step
    (agent) for 30 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde

    Mr. Malcolm Lim

    Verified customer

    Submitted Jul 05 2017

    Application
    IHC - Wholemount
    Sample
    Human Tissue (Brain)
    Specification
    Brain

    Abcam user community

    Verified customer

    Submitted Feb 22 2013

    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (sperm)
    Total protein in input
    1e+008 cells
    Specification
    sperm
    Immuno-precipitation step
    Protein A/G

    Dr. Yael Jaldety

    Verified customer

    Submitted Jun 24 2011

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