Overview

  • Product nameAnti-EGFR antibody [EP38Y]See all EGFR primary antibodies ...
  • Description
    Rabbit monoclonal [EP38Y] to EGFR
  • SpecificityThis antibody detects both EGFR phosphorylated on Tyrosine 1068 of the mature human isoform 1 (corresponding to Y1092 from the precursor form P00533-1/p170), and also unphosphorylated EGFR.
  • Tested applicationsWB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phospho-peptide corresponding to residues surrounding Tyr1068 of human EGFR.

  • Positive control
    • HeLa cell lysate or human lymph node metastasis tissue.
  • General notes

    Produced under U.S. Patent No. 5,675,063. A 40 μl trial size is available to purchase for this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52894 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
WB 1/100000. Detects a band of approximately 175 kDa (predicted molecular weight: 134 kDa).
IP 1/50.
Flow Cyt 1/100.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.

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
    • 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
    • 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 ErbB-1 antibody
    • Urogastrone antibody
    • wa2 antibody
    • Wa5 antibody
    see all

Anti-EGFR antibody [EP38Y] images

  • Overlay histogram showing A431 cells stained with ab52894 (red line). The cells were fixed with 80% 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 (ab52894, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Anti-EGFR antibody [EP38Y] (ab52894) at 1/100000 dilution + HeLa cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled. at 1/2000 dilution

    Predicted band size : 134 kDa
    Observed band size : 175 kDa (why is the actual band size different from the predicted?)
  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Ab52894 (1:100) staining human EGFR in human lymph node metastasis tissue by immunohistochemistry using paraffin embedded tissue.Ab52894 (1:100) staining human EGFR in human lymph node metastasis tissue by immunohistochemistry using paraffin embedded tissue.
  • ab52894 showing positive staining in Cervical carcinoma tissue.

  • ab52894 showing positive staining in Endometrial carcinoma tissue.

  • ab52894 showing positive staining in Glioma tissue.

  • ab52894 showing positive staining in Normal tonsil squamous cells tissue.

  • ab52894 showing positive staining in Renal cell carcinoma tissue.

References for Anti-EGFR antibody [EP38Y] (ab52894)

This product has been referenced in:
  • Brand TM  et al. Mapping C-terminal transactivation domains of the nuclear HER family receptor tyrosine kinase HER3. PLoS One 8:e71518 (2013). WB ; Human . Read more (PubMed: 23951180) »
  • Sollome JJ  et al. HER2/HER3 regulates extracellular acidification and cell migration through MTK1 (MEKK4). Cell Signal 26:70-82 (2013). Human . Read more (PubMed: 24036211) »

See all 10 Publications for this product

Product Wall


The pretreatment note in the protocol of the IHC kit refers to the antigen retrieval step that is required for most antibody to achieve a successful IHC-P staining.
For both of these antibodies (ab14440 and ab52894) I can recommend to use Sod...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100 in PBS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 25 2014



The sequence we used to raise ab52894 maps to a region surrounding Tyrosine 1068 of EGFR of human origin (Swiss Prot: P00533). While some antibodies raised against phosphorylated regions will be specific for the phosphorylated form of the pro...

Read More

Hello! We do recommend you use an antigen retrieval step, heat mediated such as citrate pH6. The antibody you suggest, ab52894 is a good one. Another EGFR antibody with good feedback is ab9482. We do guarantee our products to work as specified on the d...

Read More

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...

Read More

Paraffin blocks are commonly cut into 4 µm thickness sections and those sections are stored in a sealed box at room temperature for no more than 6 months. We have not observed any particular rapid degradation of the EGFR antigen.

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