RabMAb

Anti-EGFR (phospho Y1092) antibody [EP774Y] (ab40815)

Overview

  • Product nameAnti-EGFR (phospho Y1092) antibody [EP774Y]See all EGFR primary antibodies ...
  • Description
    Rabbit monoclonal [EP774Y] to EGFR (phospho Y1092)
  • SpecificityThe 1092 numbering includes the 24 aa signal peptide that is cleaved during EGFR maturation (the signal peptide is 24aa long based on Swiss Prot #P00533). Essentially, the Tyrosine 1068 of mature EGFR is equivalent to Tyrosine 1092 of uncleaved EGFR.
  • Tested applicationsIHC-FoFr, WB, ICC/IF, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic phospho-peptide corresponding to residues surrounding Tyr1092 of human EGFR.

  • Positive control
  • General notesProduced under U.S. Patent No. 5,675,063.

Properties

Applications

Our Abpromise guarantee covers the use of ab40815 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-FoFr Use at an assay dependent dilution. PubMed: 18948956
WB 1/500 - 1/1000. Predicted molecular weight: 170 kDa.
ICC/IF 1/250 - 1/500.
IHC-P 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.
  • 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 (phospho Y1092) antibody [EP774Y] images

  • All lanes : Anti-EGFR (phospho Y1092) antibody [EP774Y] (ab40815) at 1/50000 dilution

    Lane 1 : A431 cell lysate with cells untreated with EGF
    Lane 2 : A431 cell lysate with cells treated with EGF


    Predicted band size : 170 kDa
    Observed band size : 170 kDa
  • Immunohistochemical staining of untreated (A) and Phosphatase- treated (B) paraffin-embedded breast adenocarcinoma tissue using ab40815.
  • Immunofluorescent staining of untreated (C) and Phosphatase- treated (D) A431 cells using ab40815.
  • ab40815 staining chondrocytes in tissue section (E13.5) of mouse limb,  by immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval using 10mM Sodium Citrate buffer (pH 6.0) for 10 min at 95°C in a pressurized chamber and blocking in 5% serum for 1 hour at 25°C. Endogenous peroxidases were quenched with 3% H2o2 for 10 minutes. The primary antibody was diluted 1/250 in NGS buffer and incubated with sample for 8 hours at 4°C. A Biotin-conjugated goat polyclonal to rabbit IgG, diluted 1/500 was used as secondary. 1XTBS/0.1% Tween-20 (TBST) gave minimum background over 1XPBS as wash buffer. 

    See Abreview

  • ab40815 staining EGFR in adult mouse skin tissue by Immunohistochemistry (paraffin embedded sections). Tissue was fixed with paraformaldehyde, then a heat mediated antigen retrieval step was performed using citric buffer pH 6.0 and blocked with 10% serum for 1 hour at 20°C followed by incubation with the primary antibody, at a 1/200 dilution, for 12 hours at 4°C. Abcam biotinylated goat anti-rabbit IgG (H+L) (ready to use) was used as the secondary antibody, catalogue number ab64256.

    See Abreview

  • ab40815 showing positive staining in Papillary carcinoma of thyroid gland tissue.

  • ab40815 showing positive staining in Glioma tissue.

  • ab40815 showing positive staining in Cervical carcinoma tissue.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-EGFR (phospho Y1092) antibody [EP774Y] (ab40815)

This product has been referenced in:
  • Lee HJ  et al. Noncovalent wild-type-sparing inhibitors of EGFR T790M. Cancer Discov 3:168-81 (2013). Read more (PubMed: 23229345) »
  • Simons AL  et al. The Justy mutant mouse strain produces a spontaneous murine model of salivary gland cancer with myoepithelial and basal cell differentiation. Lab Invest 93:711-9 (2013). Read more (PubMed: 23608756) »

See all 10 Publications for this product

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EGFR is a cell membrane protein, and the signal peptide may be cleaved off after the immature protein enters the endoplasmic reticulum. The 1092 numbering includes the 24 aa signal peptide that is cleaved during ...

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The attached reference provides some additional detail about causes of and treatments for autofluorescence. As we discussed, checking your unstained tissue as well as the no primary antibody control will help yo...

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Application Western blot
Sample Human Tissue lysate - whole (stomach cancer tissue)
Loading amount 30 µg
Specification stomach cancer tissue
Treatment 10uM cannabinoid antagonist
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Apr 08 2013

Thank you for your enquiry.

ab40815 is a tissue culture supernatant product. It is not a purified antibody. However I can say that, in general, the specific antibody concentration in a product of this type is around 0.05mg/ml.

I hop...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Intestine)
Specification Intestine
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH6
Permeabilization Yes - 0.1% Tx100 in PBS
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 20 2012

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