Anti-EGFR (phospho Y1173) antibody [9H2] (ab24912)

Overview

  • Product nameAnti-EGFR (phospho Y1173) antibody [9H2]See all EGFR primary antibodies ...
  • Description
    Mouse monoclonal [9H2] to EGFR (phospho Y1173)
  • SpecificityThis is a phosphorylation site-specific monoclonal antibody. It does not interact with the non-phosphorylated EGFR nor with unrelated Tyrosine-phosphorylated proteins.
  • Tested applicationsWB, IP, ELISA, Flow Cyt, ICC, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
  • Immunogen

    Phosphopeptide conjugate (Human).

  • EpitopeThis antibody specifically interacts with the 1170 - NAEpYLRV motif corresponding to the major autophosphorylation site of human EGFR.

Properties

Applications

Our Abpromise guarantee covers the use of ab24912 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 Use a concentration of 0.5 µg/ml. Predicted molecular weight: 134 kDa.Can be blocked with EGFR (phospho Y1173) peptide (ab87004).
IP Use 1-10µg for 106 cells.
ELISA Use a concentration of 0.05 µg/ml.
Flow Cyt Use 1µg for 106 cells.
ICC Use at an assay dependent concentration.
ICC/IF Use a concentration of 0.1 - 1 µg/ml.
IHC-Fr Use at an assay dependent concentration. PubMed: 20575069

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 (phospho Y1173) antibody [9H2] images

  • Overlay histogram showing A431 cells stained with ab24912 (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 (ab24912, 1μg/1x106 cells) 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, 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.
  • All lanes : Anti-EGFR (phospho Y1173) antibody [9H2] (ab24912) at 0.5 µg/ml

    Lane 1 : Whole cell extracts of control (co) A549 tumor cells
    Lane 2 : Whole cell extracts of EGF stimulated (EGF) A549 tumor cells
    Lane 3 : Whole cell extracts of pervanadate treated (VH) A549 tumor cells

    Lysates/proteins at 20000 cells per lane.

    Secondary
    A HRP-conjugated antibody
    developed using the ECL technique

    Predicted band size : 134 kDa


    Exposure time : 30 seconds
    Recommended blocking buffer CPPT: 0.5% (w/v) casein, 1% (w/v) PEG 4000, 1% (w/v) Polyvinylpyrrolidone (PVP), 0.1% Tween 20, 10mM Tris/HCl, pH 7.4, 150mM Sodium Chloride.

References for Anti-EGFR (phospho Y1173) antibody [9H2] (ab24912)

This product has been referenced in:
  • Bradford D  et al. Characterization of the netrin/RGMa receptor neogenin in neurogenic regions of the mouse and human adult forebrain. J Comp Neurol 518:3237-53 (2010). IHC-Fr ; Mouse . Read more (PubMed: 20575069) »
  • Lee MS  et al. ErbB-2 signaling is involved in regulating PSA secretion in androgen-independent human prostate cancer LNCaP C-81 cells. Oncogene 22:781-96 (2003). Read more (PubMed: 12569372) »

See all 2 Publications for this product

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Application Western blot
Sample Human Cell lysate - whole cell (A431 cell lysate in kinase lysis buffer and 3 X sa)
Loading amount 20 µg
Specification A431 cell lysate in kinase lysis buffer and 3 X sa
Treatment Nontreated/treated with EGFR inhibtior and then unstimulated / stimulated with EGF (50ng/ml) for 15 mins
Gel Running Conditions Reduced Denaturing (9 % SDS-PAGE, 1 hr at 175 V)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: R T°C
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Mrs. surekha pimple

Verified customer

Submitted Oct 11 2010

Thank you for your enquiry. I have been in touch my colleagues in goods out. They have detailed that this antibody is supplied lyophilized and therefore the sample that you have has not been fully lyophilised. The powder itself is pink in color. Th...

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