Anti-EGFR (phospho Y1068) antibody (ab5644)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab5644 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 1/1000. Detects a band of approximately 185 kDa.

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 Y1068) antibody images

  • Peptide Competition: Extracts prepared from NIH3T3 cells expressing EGFR were starved for 30 hours, then stimulated for 10 minutes with 30 ng/mL EGF (+), or left unstimulated (-), then resolved by SDS-PAGE on a 6% Tris-glycine gel, and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 µg/mL ab5644 antibody, following prior incubation with: no peptide (1, 2), the phosphopeptide immunogen (3, 4), or, the non phosphopeptide corresponding to the phosphopeptide immunogen (5, 6). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, demonstrating the specificity of the ab5644 antibody for this phosphorylated residue. The data also show the activation of the EGFR after stimulation with EGF.
  • All lanes : Anti-EGFR (phospho Y1068) antibody (ab5644) at 1/200 dilution

    Lane 1 : A431 whole cell lysate - not treated
    Lane 2 : A431 whole cell lysate - 100ng/ml EGF for 1 minute
    Lane 3 : A431 whole cell lysate - 100ng/ml EGF for 2.5 minutes
    Lane 4 : A431 whole cell lysate - 100ng/ml EGF for 5 minutes
    Lane 5 : A431 whole cell lysate - 100ng/ml EGF for 10 minutes
    Lane 6 : A431 whole cell lysate - 100ng/ml EGF for 20 minutes
    Lane 7 : A431 whole cell lysate - 100ng/ml EGF for 40 minutes

    Secondary
    HRP conjugated Goat anti-rabbit antibody
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 2 minutes

    This image is courtesy of an Abreview submitted by Mr Samir Nuseibeh

    See Abreview

References for Anti-EGFR (phospho Y1068) antibody (ab5644)

This product has been referenced in:
  • Ather F  et al. Dacomitinib, an irreversible Pan-ErbB inhibitor significantly abrogates growth in head and neck cancer models that exhibit low response to cetuximab. PLoS One 8:e56112 (2013). WB ; Human . Read more (PubMed: 23405260) »
  • Wilson TR  et al. Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature 487:505-9 (2012). Human . Read more (PubMed: 22763448) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (heart)
Specification heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Ms. sarah shen

Verified customer

Submitted May 12 2014

Thank you for contacting us.

To our knowledge, ab5644 has not been tested in immunohistochemistry (paraffin) however, this does not mean that it will not work in this application it just means that we have not tested it ourselves. By partici...

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Application Western blot
Sample Rat Tissue lysate - whole (Liver)
Loading amount 30 µg
Specification Liver
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 25 2012

Application Western blot
Sample Human Tissue lysate - whole (Liver)
Loading amount 50 µg
Specification Liver
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 02 2012

Application Western blot
Sample Rat Tissue lysate - whole (Liver)
Loading amount 50 µg
Specification Liver
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 08 2011

Application Western blot
Sample Human Cell lysate - whole cell (Lung)
Loading amount 70 µg
Specification Lung
Treatment EGF 100ng/ml 2.5 minutes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. Samir Nuseibeh

Verified customer

Submitted Oct 23 2006

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have been in touch with the originators of ab5638, ab5644 and ab14017 with regards the band sizes that you have detected. I have received th...

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