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    egfr-phospho-y1068-antibody-y38-ab32430.pdf

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Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
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RecombinantRabMAb

Recombinant Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)

  • Datasheet
  • SDS
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Western blot - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
  • Immunoprecipitation - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
  • Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y38] to EGFR (phospho Y1068)
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-EGFR (phospho Y1068) antibody [Y38]
    See all EGFR primary antibodies
  • Description

    Rabbit monoclonal [Y38] to EGFR (phospho Y1068)
  • Host species

    Rabbit
  • Specificity

    ab32430 recognises only EGFR phosphorylated on Tyrosine 1068 of the mature human isoform 1 (corresponding to Y1092 from the precursor form P00533-1/p170).
  • Tested applications

    Suitable for: WB, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Pig
  • Immunogen

    Synthetic peptide within Human EGFR (phospho Y1068). The exact sequence is proprietary. Residues surrounding Tyr1068 of mature human EGFR.
    Database link: P00533
    (Peptide available as ab205609)

  • Positive control

    • Purchase matching WB positive control:Recombinant human EGFR protein
    • WB: A431 cell lysate treated with EGF. ICC/IF: A431 cell lysate treated with EGF.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y38
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Signal Transduction
    • Growth Factors/Hormones
    • EGF
    • Cell Biology
    • Cell Cycle
    • Cell differentiation
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • 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
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-EGFR (phospho Y1068) antibody [Y38] - BSA and Azide free (ab171022)
    • Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [Y38] (ab237420)
    • Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [Y38] (ab237421)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant human EGFR protein (Active) (ab155639)
    • Recombinant human EGFR protein (Fc Chimera) (ab84002)
  • Related Products

    • Human EGFR ELISA Kit (pY1068) (ab126438)
    • Human EGFR (pY1068) + total EGFR ELISA Kit (ab126439)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32430 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
WB
1/1000 - 1/10000. Predicted molecular weight: 135 kDa.

A blocking peptide is available for this antibody as ab205609.

ICC/IF
1/100 - 1/250.
IP
1/30.

For unpurified format use at 1/150 dilution.

Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 135 kDa.

A blocking peptide is available for this antibody as ab205609.

ICC/IF
1/100 - 1/250.
IP
1/30.

For unpurified format use at 1/150 dilution.

Application notes
Is unsuitable for Flow Cyt.

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: 397070 Pig
    • Omim: 131550 Human
    • SwissProt: P00533 Human
    • Unigene: 488293 Human
    • Unigene: 605083 Human
    • 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

    • Western blot - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
      Western blot - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
      All lanes : Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430) at 1/1000 dilution (Purified)

      Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates
      Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 minutes whole cell lysates
      Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 minutes. Then the membrane was incubated with phosphatase

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 135 kDa
      Observed band size: 175 kDa why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
      Immunoprecipitation - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)

      ab32430 (purified) at 1/30 dilution (2ug) immunoprecipitating EGFR in A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug . A431 (Human epidermoid carcinoma epithelial cell) treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
      Lane 2 (+): ab32430 & A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32430 in A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
      For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
      Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)

      Immunocytochemistry/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling EGFR (phospho Y1068) with purified ab32430 (left) and EGFR with ab52894 (right) both at a dilution of 1/1000.

      Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      The image shows increased membrane staining after EGF (100ng/ml, 30min) treatment on A431 cells. The LP treatment decreased the increased membrane staining caused by EGF.

      ab52894 was used as a Pan control for ab32430. The results showed membrane and cytoplasmic staining on untreated, EGF and EGF + LP treated A431 cells.

    • Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
      Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (34)

    Publishing research using ab32430? Please let us know so that we can cite the reference in this datasheet.

    ab32430 has been referenced in 34 publications.

    • Yin B  et al. Epidermal growth factor/epidermal growth factor receptor moves into the osteoblasts' cell nuclei where it is involved to regulate STAT5's signaling. Biofactors 47:351-362 (2021). PubMed: 33503297
    • de Wit M  et al. Mutation and drug-specific intracellular accumulation of EGFR predict clinical responses to tyrosine kinase inhibitors. EBioMedicine 56:102796 (2020). PubMed: 32512509
    • Waszkiewicz EM  et al. Expression of Insulin-Like Growth Factor 1 (IGF-1) and Epidermal Growth Factor (EGF) Receptors and the Effect of IGF-1 and EGF on Androgen and Estrogen Release in the Myometrium of Pigs-In Vitro Study. Animals (Basel) 10:N/A (2020). PubMed: 32466279
    • Xu M  et al. Proteomic Reveals Reasons for Acquired Drug Resistance in Lung Cancer Derived Brain Metastasis Based on a Newly Established Multi-Organ Microfluidic Chip Model. Front Bioeng Biotechnol 8:612091 (2020). PubMed: 33415100
    • Xia X  et al. A novel tumor suppressor protein encoded by circular AKT3 RNA inhibits glioblastoma tumorigenicity by competing with active phosphoinositide-dependent Kinase-1. Mol Cancer 18:131 (2019). PubMed: 31470874
    View all Publications for this product

    Customer reviews and Q&As

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