製品の概要

  • 製品名Anti-EGFR (phospho Y1086) antibody [Y39]
    EGFR 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [Y39] to EGFR (phospho Y1086)
  • 特異性This antibody only detects EGFR phosphorylated on Tyrosine 1086 of the mature human isoform 1.
  • アプリケーション適用あり: WB, IP, ICC/IFmore details
    適用なし: Flow Cyt or IHC
  • 種交差性
    交差種: Human
  • 免疫原

    corresponding to Human EGFR.
    (Peptide available as ab179559)

  • ポジティブ・コントロール
    • A431 cells.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

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

製品の特性

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab32086 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/1000 - 1/10000. Detects a band of approximately 170 kDa (predicted molecular weight: 134 kDa).
IP 1/150.
ICC/IF 1/100 - 1/250.
  • 追加情報Is unsuitable for Flow Cyt or IHC.
  • ターゲット情報

    • 機能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.
    • 組織特異性Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
    • 関連疾患Lung cancer
      Inflammatory skin and bowel disease, neonatal, 2
    • 配列類似性Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
      Contains 1 protein kinase domain.
    • 翻訳後修飾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.
    • 細胞内局在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
    • 参照データベース
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    Anti-EGFR (phospho Y1086) antibody [Y39] 画像

    • Immunocytochemistry/ Immunofluorescence  analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR (phospho Y1086) with ab32086 at 1/100, 3 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, a AlexaFluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000, 2 μg/ml. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclear stain was DAPI (blue).

      The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.

      For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.

    • Dot blot analysis of EGFR (phospho Y1086) phospho peptide (Lane 1), EGFR non-phospho peptide (Lane 2), labeling EGFR (phospho Y1086) with ab32068 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 5 seconds.

    • All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/3000 dilution

      Lane 1 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysates
      Lane 2 : A431 (Human epidermoid carcinoma cell line) treated with 100 ng/ml EGF for 10 minutes whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 134 kDa
      Observed band size : 170 kDa (why is the actual band size different from the predicted?)


      Exposure time : 10 seconds

      Blocking and dilution buffer: 5% NFDM/TBST. 

    • Immunofluorescent analysis of EGFR (phospho Y1086) expression in A431 cell culture, using 1/100 ab32086.
    • All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/10000 dilution

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


      Predicted band size : 134 kDa
      Observed band size : 150 kDa (why is the actual band size different from the predicted?)

    Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) 使用論文

    This product has been referenced in:
    • Johansson M  et al. The soluble form of the tumor suppressor Lrig1 potently inhibits in vivo glioma growth irrespective of EGF receptor status. Neuro Oncol 15:1200-11 (2013). WB ; Human . Read more (PubMed: 23723255) »
    • Xu ZH  et al. RAF1-MEK1-ERK/AKT axis may confer NSCLC cell lines resistance to erlotinib. Int J Clin Exp Pathol 6:1493-504 (2013). WB ; Human . Read more (PubMed: 23923067) »

    See all 6 Publications for this product

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