Anti-EGFR antibody [EGFR1] (ab30)

Reviews (4)Q&A (15)References (18)

Overview

  • Product name
    Anti-EGFR antibody [EGFR1]
    See all EGFR primary antibodies
  • Description
    Mouse monoclonal [EGFR1] to EGFR
  • Host species
    Mouse
  • Specificity

    Recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR. This antibody does not react with erbB-2, erbB-3 and erbB-4.

  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, IP, Flow Cyt, IHC - Wholemountmore details
    Unsuitable for: ELISA or WB
  • Species reactivity
    Reacts with: Mouse, Horse, Human
  • Immunogen

    Tissue, cells or virus corresponding to Human EGFR (extracellular). Human epidermoid carcinoma line A431; epitope mapped between aa 6-273 of human EGFR.

  • Epitope
    Extracellular.
  • Positive control
    • IHC-Fr: Frozen normal human placenta ICC/IF: A431 cells Flow Cyt: A 431 cells
  • General notes

    Recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR.

    This antibody clone is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab30 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
IHC-Fr Use a concentration of 10 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
Flow Cyt Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
IHC - Wholemount Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ELISA or WB.

    • Target

    • 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
    • Database links
      see all
    • Alternative names
      • 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

    Images

    • Flow Cytometry - Anti-EGFR antibody [EGFR1] (ab30)
      Flow Cytometry - Anti-EGFR antibody [EGFR1] (ab30)

      Overlay histograms showing left positive A431 cells and right negative Jurkat cells stained with ab30 (red line). The cells were incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab30, 1x106 cells in 100ul at 1ug/ml) for 30 min on ice. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) preabsorbed (ab150117) at 1/2000 dilution for 30 min on ice.

      Isotype control antibody (black line) was mouse IgG2bκ (ab170192) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EGFR1] (ab30)
      Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EGFR1] (ab30)

      ab30 staining EGFR (colored green) in A431 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab30 at 5μg/ml overnight at 4ºC. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [EGFR1] (ab30)
      Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [EGFR1] (ab30)

      IHC image of EGFR staining in a section of frozen normal human placenta, fixed in 10% paraformaldehyde (10 min). Staining was performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab30, 10ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References

    This product has been referenced in:
    • Fisher SA  et al. Photo-immobilized EGF chemical gradients differentially impact breast cancer cell invasion and drug response in defined 3D hydrogels. Biomaterials 178:751-766 (2018). Flow Cyt . Read more (PubMed: 29452913) »
    • Bäck N  et al. The endocytic pathways of a secretory granule membrane protein in HEK293 cells: PAM and EGF traverse a dynamic multivesicular body network together. Eur J Cell Biol 96:407-417 (2017). Read more (PubMed: 28377049) »
    See all 18 Publications for this product

    Publishing research using ab30? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 19 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EGFR antibody [EGFR1]

     Average
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human tissue from different sites (TMA))
    Antigen retrieval step
    Other - Buffer/Enzyme Used: Citrate pH6, Tris-EDTA pH 9 and Chymotrypsin
    Permeabilization
    No
    Specification
    Human tissue from different sites (TMA)
    Blocking step
    (agent) for 10 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Formaldehyde
    Read More

    Mr. Malcolm Lim

    Verified customer

    Submitted Jul 14 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-EGFR antibody [EGFR1]

     Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Breast cancer cell lines)
    Permeabilization
    Yes - 0.5% tritonx/PBS
    Specification
    Breast cancer cell lines
    Blocking step
    (agent) for 30 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    Read More

    Mr. Malcolm Lim

    Verified customer

    Submitted Jul 05 2017

    IHC - Wholemount abreview for Anti-EGFR antibody [EGFR1]

     Good
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Human Tissue (Brain)
    Specification
    Brain
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 22 2013

    Question

    Hi,   Can you please let me know the concentration of this antibody? Anti-EGFR antibody [EP38Y] (ab52894). I am looking for an EGFR monoclonal antibody which concentration is at least or higher than 1mg/ml.  Do you have anyone match these criteria?   Appreciate your help!!

    Read More
    Answer

    Thank you for contacting us.

    Rabbit monoclonals typically have 10-100x the affinity of mouse monoclonals, as such it is often the case that the concentration of a rabbit monoclonal will be lower than a mouse monoclonal. Ab52894 does not have a concentration of near 1 mg/ml.

    We do offer two EGFR monoclonal antibodies which do have concentrations over 1mg/ml. These are ab30 and ab62. I have included the links to those products below.

    ab30:https://www.abcam.com/EGFR-antibody-EGFR1-ab30.html

    ab62:https://www.abcam.com/EGFR-antibody-F4-ab62.html

    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

    I would like to test ab30 for use in PFA fixed floating sections

    Read More
    Answer

    DISCOUNT CODE: ***
    Expiration date: November 13th, 2012

    I am very pleased to hear you would like to accept our offer and test ab30 in IHC-PFA. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for IHC-PFA and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
    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.

    Read More

    Question

    phone call: searching for monoclonal antibody for flow cytometry on rat cells

    Read More
    Answer

    Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

    Hier ist die Liste der monoklonalen Antikörper gegen EGFR, welche wir am Telefon diskutiert haben:

    ab30, unconjugated, tested in flow cytometry and in human, mouse and horse
    https://www.abcam.com/index.html?datasheet=30 (or use the following: https://www.abcam.com/index.html?datasheet=30).
    ab11401 FITC conjugated and tested in flow cytometry, predicted to rat
    https://www.abcam.com/index.html?datasheet=11401 (or use the following: https://www.abcam.com/index.html?datasheet=11401).
    ab130738, PE conjugated, tested in flow cytometry and human and horse
    https://www.abcam.com/index.html?datasheet=130738 (or use the following: https://www.abcam.com/index.html?datasheet=130738).
    ab24293, biotin conjugated, tested in flow cytometry
    https://www.abcam.com/index.html?datasheet=24293 (or use the following: https://www.abcam.com/index.html?datasheet=24293).
    ab52894 and ab32562tested against rat, not yet in flow cytometry. will need permeabilization
    https://www.abcam.com/index.html?datasheet=52894 (or use the following: https://www.abcam.com/index.html?datasheet=52894).


    Unseres Wissens nach wurde noch keinerunserer monoklonalenAntikörper gegen EGFR in Durchflusszytometrie (flow) und auf der Ratte getestet. Falls Sie selbst einen dieser Antikörper testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert vonausgewählten Antikörpersabdecken, und gegen eine erneute Bestellung eines weiteren primären Antikörpers von uns verrechnet werden.

    Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

    1.) Bestätigen Sie mir bitte, welchen Antikörper Sie testen möchten.

    2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

    3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

    4.) Testen Sie den Antikörper in auf den Rattenzellen in Durchflusszytometrie.

    5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
    Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
    https://www.abcam.com/abreviews

    6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderenprimären Antikörperbei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummerdes getesten Antikörpersbereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

    Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

    Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
    https://www.abcam.com/collaborationdiscount

    Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

    Read More

    Question

    Thank you very much for giving me many advice.
    Last year our lab bought one antibody against EGFR (ab24293)from your company. I found it didn't work on my cell line H1703 (somebody reported EGFR was expressed on the surface of H1703 cell line with Western blotting method.)
    Yesterday you guarantee the antibody (ab30) should work follow your company method. I am planning to buy it. But my boss want to know if the antibody doesn't work again, whether your company can completely refund to us? Thank you again.

    Read More
    Answer

    Thank you for your reply.

    The antibody ab30 will be covered under our Abpromise guarantee for use in ICC on human cells. If for any reason you are unsatisfied with the results, we will be happy to refund or replace it for you as long as you contact us within six months of purchase. I hope this helps, pleaselet me know if you have any additional questionsor concerns.

    Read More

    Question

    Would like to stain EGFR of live H1703 cells.

    Read More
    Answer

    Thank you for your call today and for our discussion of these EGFR antibodies.

    Please let me know if you have any further questions or would like any assistance choosing reagents for your studies.

    I hope you have a great day!

    Read More

    Immunoprecipitation abreview for Anti-EGFR antibody [EGFR1]

     Excellent
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (sperm)
    Total protein in input
    1e+008 cells
    Specification
    sperm
    Immuno-precipitation step
    Protein A/G
    Read More

    Dr. Yael Jaldety

    Verified customer

    Submitted Jun 24 2011

    Question

    BATCH NUMBER 254 ORDER NUMBER replacem DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE Recombinant human sEGFR-extracellular http://www.researchd.com/cytokines/recpt.htm PRIMARY ANTIBODY Ab30 / diluted by PBS / 1?F1000 / room temperature for 1 hour / PBST wash 3 times SECONDARY ANTIBODY PIERCE 31320 / goat / PBS / 1?F5000 / room temperature for 1 hour / PBST wash 3 times DETECTION METHOD NEOGEN #304177 K-Blue® MAX Substrate (TMB) POSITIVE AND NEGATIVE CONTROLS USED the sample is a postive control ANTIBODY STORAGE CONDITIONS 10 ul aliquots in - 20 ?? TYPE OF ELISA Direct ELISA COATING WELL PBS 100 ng / well BLOCKING CONDITIONS StartingBlock Blocking Buffer / room temperature for 1 hour http://www.piercenet.com/Objects/View.cfm?type=ProductFamily&ID=65B71909-7ABF-4D52-8401-3107D9977640 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? none ADDITIONAL NOTES This lot is an replacement. (Enquiry detail for enquiry ID: 358940). Our customer repeated the ELISA assay three times but the signal remains to be very weak.(Raw data is available) Our customer has generated EGFR antibody which works well in ELISA and WB and needed a commercial antibody (ab30)to serve as a positive control. But it seems like the titer of ab30 is extremely low. Our customer would like to try ab231 if a replacement can be processed again for this case.

    Read More
    Answer

    I'm sorry to hear you are still having problems with the replacement of ab30, this must be very fustrating. I understand that my colleague Danielle has raised the possibility that "this antibody does not recognise recombinant EGFR. The epitope has been mapped to between aa 6-273 of human EGFR and this stretch of AAs is included in the recombinant protein used in this research. The epitope maps to the external EGF-binding domain of the EGFR transmembrane glycoprotein and perhaps this region of the recominant protein is not in the correct configuration for the antibody to bind" (e-mail from 28th Sept 2004). I would like to suggest trying an overnight incubation with the primary antibody (at 4C) to enable better binding and if this is still unsuccessful we can arrange for a credit note or replacement with ab231 (please note this antibody had not been tested for ELISA) . Please let me know if this helps and do not hesitate to contact us for further advice,

    Read More

    1-10 of 19 Abreviews or Q&A

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