Overview

  • Product nameAnti-EGFR antibody [F4]See all EGFR primary antibodies ...
  • Description
    Mouse monoclonal [F4] to EGFR
  • SpecificityNo reaction with erbB-2, erbB-3 and erbB-4.
  • Tested applicationsFlow Cyt, Sandwich ELISA, IP, ELISA, IHC-P, RIA, IHC-Frmore details
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse, Rat
  • Immunogen

    Synthetic peptide:

    DVVDADEYLIPQ

    from Human EGFR

  • EpitopeThe epitope is aa 985-996 in the cytoplasmic domain of human EGFR. Reacts with both phosphorylated and unphosphorylated EGFR.
  • Positive control
    • This antibody gave a positive result in IHC in the following FFPE tissue: Human normal placenta.

Properties

Applications

Our Abpromise guarantee covers the use of ab62 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
Flow Cyt 1/50.
Sandwich ELISA Use a concentration of 5 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to EGFR (ab2430). For sandwich ELISA, use this antibody as Capture at 5µg/ml with ab2430 as Detection.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
RIA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

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 [F4] images

  • Standard Curve for EGFR (Analyte: ab61264)dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [F4] to EGFR (ab62) at 5ug/ml and Detector Antibody Rabbit polyclonal to EGFR (ab2430) at 0.5ug/ml
  • Overlay histogram showing A431 cells stained with ab62 (red line). The cells were fixed with 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 (ab62, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • IHC image of EGFR staining in Human normal placenta formalin fixed paraffin embedded tissue section, 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 20 mins. The section was then incubated with ab62, 10µg/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. 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 for Anti-EGFR antibody [F4] (ab62)

This product has been referenced in:
  • Jaganathan S  et al. A functional nuclear epidermal growth factor receptor, SRC and stat3 heteromeric complex in pancreatic cancer cells. PLoS One 6:e19605 (2011). Human . Read more (PubMed: 21573184) »
  • Jing MX  et al. Estrogen receptor-alpha promoter methylation in sporadic basal-like breast cancer of Chinese women. Tumour Biol 32:713-9 (2011). IHC-P ; Human . Read more (PubMed: 21625942) »

See all 8 Publications for this product

Product Wall

Here is a more complete description of the EGFR fragment ab68104:

The EGFR contains the extracellular domain of the human EGFR (25-647 a.a.) excluding the signal peptide.

So, ab2430 and ab62 are not appropriate choices for detecti...

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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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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry ab62 did not perform as stated on the datasheet and for the inconvenience this has caused. As req...

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Thank you for filling in the questionnaire and for reporting this problem to us. We will use the details you sent about your protocol in order to investigate the source of the problem. I have looked through your protocol, which looks fine to me. I appr...

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Thank you for your reply. I'm assuming the EGFR will be taken from human cancer cells? This does sound similar to a sandwich ELISA. For sandwich ELISA, we would usually recommend using a polyclonal to pull down as much of the protein a...

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Thank you for contacting us. I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. I am atta...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Tumor)
Specification Tumor
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: DAKO target retrieval solution
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 08 2010

We do not have any reports of its use in FACS yet.

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