Anti-EGFR 抗体 [ICR10] (ab231)

Reviews (5)Q&A (16)References (19)

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, ELISA, ICC/IF, IP, IHC-Fr, ICCmore 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.

 
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
  • 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

    • Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [ICR10] (ab231)
      Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [ICR10] (ab231)Image courtesy of an anonymous Abreview.
      ab231 staining EGFR in human tumor cell line xenograft in mouse by Immunohistochemistry (Frozen sections).
      Tissue was fixed in acetone. Samples incubated with ab231 at a 1/100 dilution for 1 hour at 23°C. The secondary used was an undiluted HRP conjugated goat anti-rat polyclonal.

      See Abreview

    • 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

    This product has been referenced in:

    • Gerlach JP  et al. Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells. Sci Rep 9:1469 (2019). Read more (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). Read more (PubMed: 30609526) »
    See all 19 Publications for this product

    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。

    レビューと Q&A

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    1-10 of 21 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. Thanks!

    Read More
    Answer

    These four antibodies recognize the extracellular domain of EGFR and are validated for flow cytometry.

    ab231 (clone ICR10)
    ab30 (clone EGFR1)
    ab93051 (clone 31G7)
    ab32198 (clone E234)

    Clones ICR10 and EGFR1 are available directly conjugated. We do not have data demonstrating the effects of the antibodies on binding of EGF to EGFR, except for ab231, clone ICR10, 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. We do not have clone ICR12, which appears to neither block or enhance (for example, ICR9) binding of the ligand EGF to the receptor.

    Read More

    Question

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

    Read More
    Answer

    Thank you for your inquiry regarding ab231.
    We do not measure the isoelectric point of our antibodies. Most monoclonal antibodies will have an isoelectric point around 6.1, but this is not something that we test.
    Please let me know if you have any additional questions.
    .

    Read More

    Question

    Thank you. Do you have any idea when this will arrive?

    Read More
    Answer

    Thank you very much for your reply.

    I've looked at the package'stracking information, and it arrived on September 20 at 9:34 EST. You can track the package on FedEx.com with tracking number: ***. It was signed for by ***- does that help you track it down?

    Please let me know if you can find thepackage with this information, or if you need any furtherassistance and I'll be happy to help.

    Read More

    Question

    For my experiments, I am using mouse osteoblasts which express EGFR. Also, I use 2% mouse serum for blocking and a dilution of 1:100 and 1:400. Under these conditions, the secondary antibody alone is giving a positive signal. If possible, I would like to get a replacement with AB6115 as noted below. My PO number is shown below.


    Thank you for the technical support,

    Read More
    Answer

    Thank you very much for your reply.

    It does sound like the secondary is reacting with the IgG's in the mouse serum. I'm sending a free of charge vial of ab6115 on the order *** which should arrive tomorrow.

    Please keep me updated about the results using this secondary, and if there is anything else that we can do for you please let me know.

    Read More

    Question

    Inquiry: I am trying to use ab231 and ab7100 for detecting EGFR in mouse by flow cytometry. However, the secondary antibody is giving a high background signal. I am assuming this is because it reacts with mouse. Would you agree with this interpretation and if so is there another secondary antibody that you would recommend? I had previously contacted customer service and they suggested that I use ab7100. Thanks,

    Read More
    Answer

    Thank you very much for contacting us with your question.

    It is possible that the secondary antibody is causing some background signal, as ab7100 has been pre-adsorbed only against human serum proteins and not mouse serum proteins. Have you determined that the secondary antibody is causing the background due to signal in a "no primary" or isotype control sample? What dilution of ab7100 are you using, and what kind of mouse cells are you analyzing? What kind of blocking solution do you use?

    If you know that the background is coming from the secondary antibody, I'd suggest using a secondary antibody that has been pre-adsorbed against mouse serum proteins. Ab6115 is a goat anti-rat antibody that has been pre-adsorbed against mouse. I'd be happy to send this as a replacement for ab7100 if it is not working as it should be. Please send me your order or PO number if you would like to receive this replacement.

    I look forward to hearing from you. If there is anything else that we can do for you, please let me know and I'll be happy to help.

    Read More

    Question

    I would like to use the anti-EGFR antibody (ab231) for flow cytometry in the mouse. Can you recommend a secondary antibody labeled with a fluorophore?

    Read More
    Answer

    Thank you for contacting us.

    A common fluorophore used is FITC (green), and we carry ab7100 a rabbit polyclonal secondary antibody to rat iGG - H&L (FITC), pre-adsorbed as ab7100. This has been tested in FC.

    https://www.abcam.com/Rabbit-polyclonal-Secondary-Antibody-to-Rat-IgG-H-L-FITC-pre-adsorbed-ab7100.html

    If you require a different fluorophore, let me know and I will search again.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Question

    Hi,


    I was wondering how I can find that whether the following anti EGFR products :







    https://www.abcam.com/EGFR-antibody-EGFR1-Biotin-ab24293.html

    https://www.abcam.com/EGFR-antibody-EGFR1-Biotin-ab24293.html



    https://www.abcam.com/EGFR-antibody-F4-Biotin-ab24284.html

    https://www.abcam.com/EGFR-antibody-F4-Biotin-ab24284.html




    have inhibitory effect to block EGF binding or not. I would appreciate it if you could help me about it.

    Kind Regards,

    Read More
    Answer

    Thank you for your enquiry and your interest in our products.

    I can confirm that neither of these antibodies have been characterized for inhibition studies. ab24284 is suitable for IHC-P only; whilst ab24293 has been tested for Flow cytometry and IHC-Fr.

    I have conducted a search for you and found another product in our catalogue ab231 which has been tested for inhibition studies. I would advise you to take a look at the on-line datasheet to see if it suits your need:

    https://www.abcam.com/egfr-antibody-icr10-ab231.html

    I hope this helps and if I can assist further, please do not hesitate to contact me.

    Read More

    1-10 of 21 Abreviews or Q&A

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