Overview

  • Product name

    Anti-EGFR Affibody® Molecule FITC
    See all EGFR affibody® molecule
  • Conjugation

    FITC. Ex: 493nm, Em: 528nm
  • Conjugation notes

    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.
  • Specificity

    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.
  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Other Immunogen Type corresponding to EGFR.

  • Positive control

    A431 cells.
  • General notes

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

Properties

  • Form

    Liquid
  • Storage instructions

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

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

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

  • 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
  • Alternative names

    • 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
  • Database links

Applications

Our Abpromise guarantee covers the use of ab81872 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
Flow Cyt Use a concentration of 0.1 - 5 µg/ml.
ICC/IF Use a concentration of 1 - 5 µg/ml.
  • Application notes
    Is unsuitable for IHC-P.
  • Images

    • 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).
    • 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).
    • 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.

    References

    This product has been referenced in:

    • 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). Read more (PubMed: 26330166) »
    See 1 Publication for this product

    Customer reviews and Q&As

    1-7 of 7 Q&A

    Answer

    Thank you for providing that information.

    I will go ahead and upload the information that you provided. It may take a week or so for it to appear on the webpages.

    Once again, thank you for providing that information and for your patience in this matter.

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    Question
    Answer

    Thank you for your call today, and I'm sorry for the trouble with submitting your review.

    Please go ahead and send the protocol information and any data that you have on Ab81872, and I will upload the review manually, as we currently don't have a review form for Affibody products.

    I look forward to hearing from you and receiving the review. Please let me know if you have any questions or if there is anything else that we can do for you.

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    Question
    Answer

    Thank you for your call today, and I'm sorry for the trouble with submitting your review.

    As we discussed, please go ahead and send the protocol information and any data that you have on Ab81872, and I will upload the review manually, as we currently don't have a review form for Affibody products.

    I look forward to hearing from you and receiving the review. Please let me know if you have any questions or if there is anything else that we can do for you.

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    Answer

    DISCOUNT CODE: *******Expiration date: ****** I am very pleased to hear you would like to accept our offer and test ab81872 inmouse tissue. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse tissue and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.  

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    Question
    Answer

    Thank you very much for your interest in ab81872. To our knowledge, ab81872 has not been tested in mouse tissue. Therefore, I can offer a discount off a future purchase if you buy ab81872 now, test it in mouse tissue and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab81872 in mouse tissue. I will then send a discount code. This code must be issued before purchasing ab81872 so please wait for my reply before ordering. 2. Purchase ab81872 either by phone, fax, or online (www.abcam.com). 3. Test it in mouse tissue. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab81872  turns out to be unsuitable for mouse tissue, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.  

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    Question
    Answer

    Thanks for your inquiry. 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] Please feel free to contact us if you require further assistance.

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    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.

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