Anti-EGFR (phospho S1070 + S1071) antibody [EP2259Y] (ab76300)

Overview

  • Product name
    Anti-EGFR (phospho S1070 + S1071) antibody [EP2259Y]
    See all EGFR primary antibodies
  • Description
    Rabbit monoclonal [EP2259Y] to EGFR (phospho S1070 + S1071)
  • Specificity
    The phosphorylation site of S1070/S1071 numbering includes the 24aa signal peptides which are removed in the mature form of EGFR and is equal to S1046/1047.
  • Tested applications
    Suitable for: WB, IP, ICC/IFmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A synthetic phospho-peptide corresponding to residues surrounding Serine 1070 and 1071 of human EGF receptor.

  • Positive control
    • WB: A431 cell lysates treated with EGF. WB: A431 cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Applications

Our Abpromise guarantee covers the use of ab76300 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 1/500000 - 1/1e+006. Detects a band of approximately 150 kDa (predicted molecular weight: 134 kDa).
IP 1/30.
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for Flow Cyt or IHC-P.
  • Target

    • Function
      Receptor 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 specificity
      Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
    • Involvement in disease
      Lung cancer
      Inflammatory skin and bowel disease, neonatal, 2
    • Sequence similarities
      Belongs 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. Dephosphorylated by PTPN1 and PTPN2.
      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 occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
      Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.
    • Cellular localization
      Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF).
    • 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
      • erb-b2 receptor tyrosine kinase 1 antibody
      • ERBB antibody
      • ERBB1 antibody
      • Errp antibody
      • HER1 antibody
      • mENA antibody
      • NISBD2 antibody
      • Oncogen ERBB antibody
      • PIG61 antibody
      • Proto-oncogene c-ErbB-1 antibody
      • Receptor tyrosine protein kinase ErbB 1 antibody
      • Receptor tyrosine-protein kinase ErbB-1 antibody
      • SA7 antibody
      • Species antigen 7 antibody
      • Urogastrone antibody
      • v-erb-b Avian erythroblastic leukemia viral oncogen homolog antibody
      • wa2 antibody
      • Wa5 antibody
      see all

    Images

    • All lanes : Anti-EGFR (phospho S1070 + S1071) antibody [EP2259Y] (ab76300) at 1/1000000 dilution

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

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 134 kDa
      Observed band size : 180 kDa (why is the actual band size different from the predicted?)
    • Immunocytochemistry/Immunofluorescence analysis of untreated and EGF (100ng/mL for 10 minutes) treated A431 cells labelling EGFR (phospho S1070 + S1071) with ab76300 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889, an Alexa Fluor® 594-conjugated mouse anti-tubulin (1/200). Nuclei counterstained with DAPI (blue).

      Control 1: untreated A431 cells incubated with PBS instead of primary antibody followed by incubation with ab150077 Alexa Fluor® 488 goat anti-rabbit IgG.
      Control 2: EGF treated A431 cells incubated with PBS instead of primary antibody followed by incubation with ab150077 Alexa Fluor® 488 goat anti-rabbit IgG.

    • Primary antibody: ab76300 at a dilution of 1/1000.

      Secondary antibody: Peroxidase conjugated-goat anti-rabbit IgG, (H+L) at a dilution of 1/2500.

      Blocking and dilution buffer: 5% NFDM/TBST

      Lane 1: EGFR (pS1070/pS1071) phospho peptide.
      Lane 2: EGFR non-phospho peptide.

      Exposure time: 3 minutes.

    References

    This product has been referenced in:
    • Singh RK & Lokeshwar BL The IL-8-regulated chemokine receptor CXCR7 stimulates EGFR signaling to promote prostate cancer growth. Cancer Res 71:3268-77 (2011). Read more (PubMed: 21398406) »

    See 1 Publication for this product

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

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