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Anti-EGFR Affibody® Molecule FITC (ab81872)

Q&A (2)References (5)
Flow Cytometry - Anti-EGFR Affibody® Molecule FITC (ab81872)
  • Flow Cytometry - Anti-EGFR Affibody® Molecule FITC (ab81872)
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR Affibody® Molecule FITC (ab81872)

製品の概要

  • 製品名

    Anti-EGFR Affibody® Molecule FITC

  • 標識

    FITC. Ex: 493nm, Em: 528nm

  • 特記事項（標識）

    The molecule is conjugated at the unique C-terminal cysteine using a maleimide activated fluorescein reagent. Stability: There is no decrease in performance of the fluorescein conjugated Affibody® molecule after 10 repeated freeze and thaw cycles or after storage for 1 week in room temperature.

  • 特異性

    This product binds to human EGFR. This anti-EGFR Affibody® molecule is a specific affinity ligand selected against the extracellular domain of EGFR. The FITC conjugated Anti-EGFR Affibody® molecule is excellent as a one step reagent for fluorescence studies of EGFR expression on cells as well as for flow cytometry.

  • アプリケーション

    適用あり: Flow Cyt, ICC/IFmore details
    適用なし: IHC-P

  • 種交差性

    交差種: Human

  • 免疫原

    Recombinant full length protein corresponding to EGFR.

  • ポジティブ･コントロール

    A431 cells.

  • 特記事項

    This product is a recombinant protein produced in E.coli.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    pH: 7.20
    Preservative: 0.02% Sodium azide
    Constituents: 0.87% Sodium chloride, 0.328% Sodium phosphate
  • Concentration information loading...

  • 特記事項（精製）

    Purity: >98% as determined by RP-HPLC analysis. Extinction coefficient: 1 Abs280 = 1.30 mg/ml.

  • 研究分野

  • 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

  • 別名

    • Avian erythroblastic leukemia viral (v erb b) oncogene homolog
    • Cell growth inhibiting protein 40
    • Cell proliferation inducing protein 61
    • EGF R
    • EGFR
    • EGFR_HUMAN
    • Epidermal growth factor receptor
    • Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog)
    • Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian)
    • erb-b2 receptor tyrosine kinase 1
    • ERBB
    • ERBB1
    • Errp
    • HER1
    • mENA
    • NISBD2
    • Oncogen ERBB
    • PIG61
    • Proto-oncogene c-ErbB-1
    • Receptor tyrosine protein kinase ErbB 1
    • Receptor tyrosine-protein kinase ErbB-1
    • SA7
    • Species antigen 7
    • Urogastrone
    • v-erb-b Avian erythroblastic leukemia viral oncogen homolog
    • wa2
    • Wa5
    see all

  • 参照データベース

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab81872の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    Flow Cyt
    Use a concentration of 0.1 - 5 µg/ml.
    ICC/IF
    Use a concentration of 1 - 5 µg/ml.
    特記事項
    Flow Cyt
    Use a concentration of 0.1 - 5 µg/ml.
    ICC/IF
    Use a concentration of 1 - 5 µg/ml.
    追加情報
    Is unsuitable for IHC-P.

    画像

    • Flow Cytometry - Anti-EGFR Affibody® Molecule FITC (ab81872)
      Flow Cytometry - Anti-EGFR Affibody® Molecule FITC (ab81872)
      Fluorescence histogram from flow cytometry analysis on EGFR expressing A431 cells. Staining with ab81872 caused an increase in fluorescence intensity as shown by a shift to the right (red line) compared to control (black line).
    • Flow Cytometry - Anti-EGFR Affibody® Molecule FITC (ab81872)
      Flow Cytometry - Anti-EGFR Affibody® Molecule FITC (ab81872)
      Titration diagram from cytometry analysis of EGFR expressing A431 cells. ab81872 was titrated on EGFR expressing A431 cells. The concentration was plotted against the mean fluorescence intensity (MFI).
    • Immunocytochemistry/ Immunofluorescence - Anti-EGFR Affibody® Molecule FITC (ab81872)
      Immunocytochemistry/ Immunofluorescence - Anti-EGFR Affibody® Molecule FITC (ab81872)
      A431 cells were stained with ab81872 at 2µg/ml by Immunofluorescence. The staining was localized to the membrane of the EGFR-expressing A431 cells. The nuclei were counter stained with DAPI.

    参考文献 (5)

    ab81872 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。

    ab81872 は 5 報の論文で使用されています。

    • Zhang S  et al. Multivalent Ligand-Nanoparticle Conjugates Amplify Reactive Oxygen Species Second Messenger Generation and Enhance Epidermal Growth Factor Receptor Phosphorylation. Bioconjug Chem 33:1716-1728 (2022). PubMed: 35993676
    • Manzano S  et al. C3G downregulation induces the acquisition of a mesenchymal phenotype that enhances aggressiveness of glioblastoma cells. Cell Death Dis 12:348 (2021). PubMed: 33824275
    • Liu YL  et al. Spatial EGFR Dynamics and Metastatic Phenotypes Modulated by Upregulated EphB2 and Src Pathways in Advanced Prostate Cancer. Cancers (Basel) 11:N/A (2019). PubMed: 31805710
    • Liu X  et al. Affinity-Tuned ErbB2 or EGFR Chimeric Antigen Receptor T Cells Exhibit an Increased Therapeutic Index against Tumors in Mice. Cancer Res 75:3596-607 (2015). PubMed: 26330166
    • McCluskey AJ  et al. Targeting HER2-positive cancer cells with receptor-redirected anthrax protective antigen. Mol Oncol 7:440-51 (2013). PubMed: 23290417

    レビューと Q&A

    Q&A
    質問を送る

    1-2 of 2 Q&A

    Question

    Do you know if this antibody is neutralizing/blocks ligand binding?

    Read More
    Answer

    ab81872 does neutralize or block ligand binding. See PubMed reference below:Nucl Med Biol. 2007 Aug;34(6):609-18. From the abstract of that paper: "The cellular binding and processing of an epidermal growth factor receptor (EGFR) targeting affibody molecule, (Z(EGFR:955))(2), was studied. This new and small molecule is aimed for applications in nuclear medicine. The natural ligand epidermal growth factor (EGF) and the antibody cetuximab were studied for comparison.METHODS: All experiments were made with cultured A431 squamous carcinoma cells. Receptor specificity, binding time patterns, retention and preliminary receptor binding site localization studies were all made after (125)I labeling. Internalization was studied using Oregon Green 488, Alexa Fluor 488 and CypHer5E markers.RESULTS: [(125)I](Z(EGFR:955))(2) and [(125)I]cetuximab gave a maximum cellular uptake of (125)I within 4 to 8 h of incubation, while [(125)I]EGF gave a maximum uptake already after 2 h. The retention studies showed that the cell-associated fraction of (125)I after 48 h of incubation was approximately 20% when delivered as [(125)I](Z(EGFR:955))(2) and approximately 25% when delivered as [(125)I]cetuximab. [(125)I]EGF-mediated delivery gave a faster (125)I release, where almost all cell-associated radioactivity had disappeared within 24 h. All three substances were internalized as demonstrated with confocal microscopy. Competitive binding studies showed that both EGF and cetuximab inhibited binding of (Z(EGFR:955))(2) and indicated that the three substances competed for an overlapping binding site.CONCLUSION: The results gave information on cellular processing of radionuclides when delivered with (Z(EGFR:955))(2) in comparison to delivery with EGF and cetuximab. Competition assays suggested that [(125)I](Z(EGFR:955))(2) bind to Domain III of EGFR. The affibody molecule (Z(EGFR:955))(2) can be a candidate for EGFR imaging applications in nuclear medicine.PMID:17707800[PubMed - indexed for MEDLINE]

    Read More

    Question

    This is regarding the FITC conjugated antibody against EGF receptor. What is the monoclonal that was used for this product? Do you know what epitope it is directed against?

    Read More
    Answer

    Thank you for contacting us. Please be advised that ab81872 is not a conjugated antibody it is in fact an Affibody molecules. These are a novel class of antibody mimetics with superior characteristics surpassing monoclonal antibodies (mAbs) and antibody fragments. Affibody® molecules are only 6 kDa in size, have an inert format (no Fc function), and have successfully been tested in humans as targeting moiety. The ab81872 is specific to the extracellular domain of EGF receptor. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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