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Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
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Anti-EGFR antibody [EGFR1] (ab30)

  • Datasheet
  • SDS
Reviews (4)Q&A (4)References (27)

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Flow Cytometry - Anti-EGFR antibody [EGFR1] (ab30)
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EGFR1] (ab30)
  • Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [EGFR1] (ab30)

Key features and details

  • Mouse monoclonal [EGFR1] to EGFR
  • Suitable for: IHC-Fr, ICC/IF, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG2b

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Overview

  • Product name

    Anti-EGFR antibody [EGFR1]
    See all EGFR primary antibodies
  • Description

    Mouse monoclonal [EGFR1] to EGFR
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-Fr, ICC/IF, Flow Cytmore details
    Unsuitable for: ELISA or WB
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Horse
  • 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: A431 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    EGFR1
  • Myeloma

    P3-NS1/1-Ag4-1
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Signal Transduction
    • Growth Factors/Hormones
    • EGF
    • Cell Biology
    • Cell Cycle
    • Cell differentiation
    • Cancer
    • Growth factors
    • EGF
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • Receptor tyrosine kinases
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Growth factor receptors
    • Cancer
    • Tumor biomarkers
    • Receptors

Associated products

  • Affibody® Molecule

    • Anti-EGFR Affibody® Molecule Agarose Immobilized (ab36039)
  • Alternative Versions

    • PE Anti-EGFR antibody [EGFR1] (ab130738)
    • PE/Cy7® Anti-EGFR antibody [EGFR1] (ab239309)
    • Biotin Anti-EGFR antibody [EGFR1] (ab24293)
    • Anti-EGFR antibody [EGFR1] - BSA and Azide free (ab264523)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

    • Recombinant human EGFR protein (Fc Chimera) (ab84002)
  • Related Products

    • Human EGFR ELISA Kit (ERBB) (ab100505)
    • Human soluble EGFR ELISA Kit (ab193764)

Applications

The Abpromise guarantee

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 (1)
Use a concentration of 5 µg/ml.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Notes
IHC-Fr
Use a concentration of 10 µg/ml.
ICC/IF
Use a concentration of 5 µg/ml.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

  • 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).
    • Target information above from: UniProt accession P00533 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 1956 Human
      • Entrez Gene: 13649 Mouse
      • Omim: 131550 Human
      • SwissProt: P00533 Human
      • SwissProt: Q01279 Mouse
      • Unigene: 488293 Human
      • Unigene: 605083 Human
      • Unigene: 420648 Mouse
      • Unigene: 439882 Mouse
      • Unigene: 8534 Mouse
      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.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (27)

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

    ab30 has been referenced in 27 publications.

    • Lima Moura S  et al. Matrix Effect in the Isolation of Breast Cancer-Derived Nanovesicles by Immunomagnetic Separation and Electrochemical Immunosensing-A Comparative Study. Sensors (Basel) 20:N/A (2020). PubMed: 32054015
    • Zhao L  et al. SGCE Promotes Breast Cancer Stem Cells by Stabilizing EGFR. Adv Sci (Weinh) 7:1903700 (2020). PubMed: 32714745
    • Wu QQ  et al. The 5-Lipoxygenase Inhibitor Zileuton Protects Pressure Overload-Induced Cardiac Remodeling via Activating PPARa. Oxid Med Cell Longev 2019:7536803 (2019). PubMed: 31781348
    • Liu C  et al. Enhanced blood-brain-barrier penetrability and tumor-targeting efficiency by peptide-functionalized poly(amidoamine) dendrimer for the therapy of gliomas. Nanotheranostics 3:311-330 (2019). PubMed: 31687320
    • Kim CS  et al. Leader cell PLC?1 activation during keratinocyte collective migration is induced by EGFR localization and clustering. Bioeng Transl Med 4:e10138 (2019). PubMed: 31572796
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-8 of 8 Abreviews or Q&A

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

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

    Abcam user community

    Verified customer

    Submitted Feb 22 2013

    Immunoprecipitation abreview for Anti-EGFR antibody [EGFR1]

    Excellent
    Abreviews
    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

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

    Read More

    Abcam community

    Verified customer

    Asked on May 24 2004

    Answer

    Regarding ab30, EGFR1 recognises the whole receptor protein rather than a truncated version.

    Read More

    Abcam Scientific Support

    Answered on Jun 09 2004

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on May 24 2004

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on May 28 2004

    Question

    We are interested in purchasing the anti-EGFR antibody, Ab30, to use in immunohistochemistry experiments. My question relates to whether this antibody detects the mutant EGFR, EGFRvIII which has a deletion of a portion of the ECD. We are interested in antibodies which DO NOT pick up EGFRvIII.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 20 2001

    Answer

    The antibody recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein and the epitope was mapped to between aa 6-273 of human EGFR. We do not know which AA seq are deleted in EGFRvIII and therefore whether the epitope lies within the deleted region.

    Read More

    Danielle Miller

    Abcam Scientific Support

    Answered on Nov 21 2001

    Question

    you are the only purveyor of EGFR-1 (wild type vs VIII specific) that claims that it can be used on formalin-fixed, paraffin embedded tissue; all others producing it say it is not suitable for this application. Can you please tell me which antigen retrieval methods (protease, microwave, pressure cooker) you use with this Mab?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 09 2001

    Answer

    In the following references the EGFR1 antibody has been used on paraffin-embedded sections: Ioachim E, et al. (1996) Anticancer Res 16(5B):3141-7. Nakopoulou L, et al. (1994) Nephrol Dial Transplant 9(7):764-9. Gorgoulis V, et al. (1992) Anticancer Res 12(4):1183-7.

    Read More

    Abcam Scientific Support

    Answered on Feb 12 2001

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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