Overview

  • Product nameAnti-EGFR antibody [EGFR1]See all EGFR primary antibodies ...
  • Description
    Mouse monoclonal [EGFR1] to EGFR
  • SpecificityThis antibody does not react with erbB-2, erbB-3 and erbB-4.
  • Tested applicationsIHC-Fr, ICC/IF, IP, Flow Cyt, IHC - Wholemount more details
  • Species reactivity
    Reacts with: Mouse, Horse, Human
  • Immunogen

    Human epidermoid carcinoma line A431; epitope mapped between aa 6-273 of human EGFR.

  • EpitopeExtracellular.
  • General notesRecognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR.

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 Notes
IHC-Fr Use at an assay dependent dilution.
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent dilution.
Flow Cyt Use 1µg for 106 cells. (unsuitable for fixed cells)
IHC - Wholemount Use at an assay dependent concentration.
  • Application notesIs unsuitable for ELISA or 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.
    • 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 antibody [EGFR1] images

    • ICC/IF image of ab30 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 (ab30, 1µg/ml) 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.

    • Overlay histogram showing HEK293 cells stained with ab30 (red line). The cells were incubated with the antibody (ab30, 1µg/1x106 cells) for 30 min at 4°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/200 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
      Please note that 4% paraformaldehyde (10 min) or methanol (5 min) fixation prior to antibody staining significantly decreased signal.

    References for Anti-EGFR antibody [EGFR1] (ab30)

    This product has been referenced in:
    • Jaldety Y  et al. Sperm epidermal growth factor receptor (EGFR) mediates a7 acetylcholine receptor (AChR) activation to promote fertilization. J Biol Chem 287:22328-40 (2012). IP . Read more (PubMed: 22577141) »
    • Boersma YL  et al. Bispecific designed ankyrin repeat proteins (DARPins) targeting epidermal growth factor receptor inhibit A431 cell proliferation and receptor recycling. J Biol Chem 286:41273-85 (2011). Flow Cyt ; Human . Read more (PubMed: 21979953) »

    See all 4 Publications for this product

    Product Wall

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

    Abcam user community

    Verified customer

    Submitted Feb 22 2013

    Thank you for contacting us.

    Rabbit monoclonals typically have 10-100x the affinity of mouse monoclonals, as such it is often the case that the concentration of a rabbit monoclonal will be lower than a mouse monoclonal. Ab52894 does not ha...

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    DISCOUNT CODE: ***
    Expiration date: November 13th, 2012

    I am very pleased to hear you would like to accept our offer and test ab30 in IHC-PFA. This code will give you: 1 free primary antibody before the expiration date. To redeem this of...

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    Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

    Hier ist die Liste der monoklonalen Antikörper gegen EGFR, welche wir am Telefon diskutiert haben:

    ab30, unconjugated, tested in flow cytometry a...

    Read More

    Thank you for your reply.

    The antibody ab30 will be covered under our Abpromise guarantee for use in ICC on human cells. If for any reason you are unsatisfied with the results, we will be happy to refund or replace it for you as long as you c...

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    Thank you for your call today and for our discussion of these EGFR antibodies.

    Please let me know if you have any further questions or would like any assistance choosing reagents for your studies.

    I hope you have a great day!

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

    Dr. Yael Jaldety

    Verified customer

    Submitted Jun 24 2011

    I'm sorry to hear you are still having problems with the replacement of ab30, this must be very fustrating. I understand that my colleague Danielle has raised the possibility that "this antibody does not recognise recombinant EGFR. The epitope has bee...

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    The protocol that your customer is using looks fine so I would be happy to send you a replacement vial just in case there is a problem with the original vial that they received. However, it is possible that this antibody does not recognise recombinant ...

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    Thank you again for your patience. Regarding ab30, EGFR1 recognises the whole receptor protein rather than a truncated version. For ab5636, the synthetic peptide immunogen to produce the antibody is approximately 10 amino acid residues around EGFR [pY8...

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