Anti-EGFR 抗体 [ICR10] (ab231)

Reviews (5)Q&A (5)References (21)

Key features and details

  • Rat monoclonal [ICR10] to EGFR
  • Suitable for: Inhibition Assay, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Human
  • Isotype: IgG2a

製品の概要

  • 製品名

    Anti-EGFR antibody [ICR10]
    EGFR 一次抗体 製品一覧

  • 製品の詳細

    Rat monoclonal [ICR10] to EGFR

  • 由来種

    Rat

  • アプリケーション

    適用あり: Inhibition Assay, Flow Cyt, ICC/IFmore details
    適用なし: WB

  • 種交差性

    交差種: Mouse, Human

  • 免疫原

    Extracellular domain of Human EGF-receptor from head and neck carcinoma

  • エピトープ

    ICR10 binds to epitope B, and has an affinity of 6.7x10-9 M.

  • ポジティブ･コントロール

    • FACS: A431 cells IHC: Breast Carcinoma tissue WB: HN5 cell lysate

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab231 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
Inhibition Assay Use at an assay dependent concentration. See ELISA Abreview.
Flow Cyt 1/50 - 1/100.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 
ICC/IF Use a concentration of 5 µg/ml.
  • 追加情報
    Is unsuitable for WB.

    • 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

      see all

    • 別名

      • 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

    画像

    • Flow Cytometry - Anti-EGFR antibody [ICR10] (ab231)
      Flow Cytometry - Anti-EGFR antibody [ICR10] (ab231)
      Staining of A431 cells with ab231 visualised with F(ab')2 rabbit anti-rat FITC conjugated secondary.
    • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [ICR10] (ab231)
      Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [ICR10] (ab231)

      ICC/IF image of ab231 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab231, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    • Inhibition Assay - Anti-EGFR antibody [ICR10] (ab231)
      Inhibition Assay - Anti-EGFR antibody [ICR10] (ab231)

      ab231 used undiluted in inhibition assay against human recombinant protein EGFR1:Fc to inhibit binding to EGF:Biotin. The primary incubated with sample for 1 hour. The antibody inhibits the binding of biotinylated EGF to coated EGFR:Fc with an IC50 ~ 5 nM.

    参考文献

    ab231 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab231 は 21 報の論文で使用されています。

    • Gerlach JP  et al. Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells. Sci Rep 9:1469 (2019). PubMed: 30728416
    • Ortega FG  et al. EGFR detection in extracellular vesicles of breast cancer patients through immunosensor based on silica-chitosan nanoplatform. Talanta 194:243-252 (2019). PubMed: 30609526
    • Lingemann M  et al. The alpha-1 subunit of the Na+,K+-ATPase (ATP1A1) is required for macropinocytic entry of respiratory syncytial virus (RSV) in human respiratory epithelial cells. PLoS Pathog 15:e1007963 (2019). PubMed: 31381610
    • Wang H  et al. Evaluation of serum extracellular vesicles as noninvasive diagnostic markers of glioma. Theranostics 9:5347-5358 (2019). PubMed: 31410219
    • Stolarczyk M  et al. Extracellular oxidation in cystic fibrosis airway epithelium causes enhanced EGFR/ADAM17 activity. Am J Physiol Lung Cell Mol Physiol 314:L555-L568 (2018). PubMed: 29351448
    View all Publications for this product

    レビューと Q&A

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    1-10 of 10 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-EGFR antibody [ICR10]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (CB17/SCID BxPC-3 xenograft)
    Specification
    CB17/SCID BxPC-3 xenograft
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Dr. Jennifer Dembinski

    Verified customer

    投稿 May 14 2009

    Western blot abreview for Anti-EGFR antibody [ICR10]

     Good
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A431 cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Specification
    A431 cells
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Aug 16 2017

    Flow Cytometry abreview for Anti-EGFR antibody [ICR10]

     Excellent
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (HCC827 cells)
    Permeabilization
    No
    Gating Strategy
    Live cells (PI negative)
    Specification
    HCC827 cells
    Preparation
    Cell harvesting/tissue preparation method: Trypsinized
    Sample buffer: RPMI1640 with 10% FBS and Penicillin-Streptomycin
    Fixation
    No
    Read More

    Abcam user community

    Verified customer

    投稿 Jul 14 2017

    Question

    I am looking for an EGFR antibody that recognizes the N-term (extracellular domain) of EGFR but does not block EGF binding to EGFR. I need the antibody for flow cytometry experiments.

    Read More
    Answer

    This antibody recognizes the extracellular domain of EGFR and is validated for flow cytometry. There is data demonstrating the effects of the antibody on binding of EGF to EGFR in the reference at the following link.

    http://onlinelibrary.wiley.com/doi/10.1002/ijc.11055/pdf

    The data in figure 3 shows that ICR10 blocks partially, compared to the medium control. receptor.

    Read More

    Question

    Do you know the isoelectric point of the Rat monoclonal ICR10 antibody, abcam part number ab231?

    Read More
    Answer

    Most monoclonal antibodies will have an isoelectric point around 6.1, but this is not something that we test except for some of our RabMab antibodies.

    Read More

    Immunohistochemistry (Frozen sections) abreview for Anti-EGFR antibody [ICR10]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (tumor cell line xenograft in mouse)
    Specification
    tumor cell line xenograft in mouse
    Fixative
    Acetone
    Permeabilization
    No
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 24 2012

    ELISA abreview for Anti-EGFR antibody [ICR10]

     Good
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Recombinant protein (EGFR1:Fc)
    Specification
    EGFR1:Fc
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Type
    Other
    Read More

    Ms. Lakshmi Ganesan

    Verified customer

    投稿 Nov 20 2009

    Question

    Customer would like to know if these EGFR antibodies are specific for the truncated receptor.

    Read More
    Answer

    Thank you for your patience. I have received the following information thus far: Ab231: In Western blotting ab231 detects a 170kDa band which corresponds to EGFR. The epitope recognized by this antibody is located on the extracellular domain of the EGFR. I belive that the truncated form of EGFR lacks the cytoplasmic and transmembrane domain. If it is indeed this simple, then I would expect the antibody to recognise the truncated form. However, to our knowledge, this antibody has not been tested for reactivity with this form of EGFR and therefore we would be unable to offer any guarantees. Ab2430: Ab2430 is made using a peptide corresponding to AA 1005-1016 of the human EGFR precursor. There are several truncated forms of EGFR and one of the more prevelant is EGFRvIII where amino acids 6-273 are replaced by a glycine residue. This antibody will recognize this variant, but it will also recognize the wild type. The researcher will need to compare the variants sequence to that of the wild type to make sure that this antibody's epitope has not been deleted. Once I receive more information regarding ab5636 and ab30, I will contact you again.

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    Question

    I am not familiar with what epitope "B" means on the data sheet.

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    Answer

    The epitope letters label 5 distinct epitopes A-E, where A is the the nearest to the N-terminus of the receptor. The following references give more information concerning these epitopes and therefore may be of assistance to you. Modjtahedi, H et al. 1993. Br J Cancer. 67:247-253 Modjtahedi, H and Dean, CJ. 1994. Int J Oncol.4:277-296

    Read More

    Question

    I want to detect mouse circulating EGFR tuncated forms (p60 and p110) which represent the extracellular domain. Will this work?

    Read More
    Answer

    This antibody has not been tested in this application, but as the epitope is on the extracellular domain I would expect this antibody to work, although we cannot guarantee it.

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