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  1. Link

    products/primary-antibodies/erbb4--her4-phospho-y1284-antibody-epr22732-ab109273.pdf

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Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
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RecombinantRabMAb

Recombinant Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)

  • Datasheet
  • SDS
Reviews (1)Q&A (2)References (6)

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Western blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
  • Western blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
  • Dot Blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
  • Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2273(2)] to ErbB4 / HER4 (phospho Y1284)
  • Suitable for: WB, Dot blot
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Protein
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Recombinant human ErbB4 / HER4 protein (ab155610)

View more associated products

Overview

  • Product name

    Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)]
    See all ErbB4 / HER4 primary antibodies
  • Description

    Rabbit monoclonal [EPR2273(2)] to ErbB4 / HER4 (phospho Y1284)
  • Host species

    Rabbit
  • Specificity

    This antibody detects ErbB4 / HER4 phosphorylated at Tyrosine 1284 and may also detect Erbb2/HER2 phosphorylated at Tyrosine 1248.

    Stimulation may be required to allow detection of the phosphorylated protein.  Please see images below for recommended treatment conditions and positive controls.

  • Tested applications

    Suitable for: WB, Dot blotmore details
    Unsuitable for: Flow Cyt (Intra),ICC/IF,IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431 treated with 50mM pervanadate for 5 min whole cell lysate and A431 treated with 100 ng/ml Epidermal Growth Factor (EGF) for 30 min whole cell lysate.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2273(2)
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Signal Transduction
    • Growth Factors/Hormones
    • EGF
    • Cancer
    • Growth factors
    • EGF
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Growth factors
    • Cancer
    • Tumor biomarkers
    • Oncoproteins
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] - BSA and Azide free (ab232527)
  • Isotype control

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

    • A-431 nuclear extract lysate (EGF treated) (ab14635)
  • Recombinant Protein

    • Recombinant human ErbB4 / HER4 protein (ab155610)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109273 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)
1/1000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 147 kDa).
Dot blot
1/1000.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 147 kDa).
Dot blot
1/1000.
Application notes
Is unsuitable for Flow Cyt (Intra),ICC/IF,IHC-P or IP.

Target

  • Function

    Specifically binds and is activated by neuregulins, NRG-2, NRG-3, heparin-binding EGF-like growth factor, betacellulin and NTAK. Interaction with these factors induces cell differentiation. Not activated by EGF, TGF-A, and amphiregulin. The C-terminal fragment (CTF) of isoform JMA-A CYT-2 (containing E4ICD2) can stimulate transcription in the presence of YAP1. ERBB4 intracellular domain is involved in the regulation of cell growth. Conflicting reports are likely due at least in part to the opposing effects of the isoform-specific and nuclear-translocated ERBB4 intracellular domains (E4ICD1 and E4ICD2). Overexpression studies in epithelium show growth inhibition using E4ICD1 and increased proliferation using E4ICD2. E4ICD2 has greater in vitro kinase activity than E4ICD1. The kinase activity is required for the nuclear translocation of E4ICD2.
  • Tissue specificity

    Expressed at highest levels in brain, heart, kidney, in addition to skeletal muscle, parathyroid, cerebellum, pituitary, spleen, testis and breast. Lower levels in thymus, lung, salivary gland, and pancreas. Isoform JM-A CYT-1 and isoform JM-B CYT-1 are expressed in cerebellum, but only the isoform JM-B is expressed in the heart.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Isoform JM-A CYT-1 and isoform JM-A CYT-2 but not isoform JM-B CYT-1 and isoform JM-B CYT-2 are processed by ADAM17. Proteolytic processing in response to ligand or 12-O-tetradecanoylphorbol-13-acetate stimulation results in the production of 120 kDa soluble receptor forms and intermediate membrane-anchored 80 kDa fragments (m80HER4), which are further processed by a presenilin-dependent gamma-secretase to release the respective cytoplasmic intracellular domain E4ICD (either E4ICD1/s80Cyt1 or E4ICD2/s80Cyt2). Membrane-anchored 80 kDa fragments of the processed isoform JM-A CYT-1 are more readily degraded by the proteasome than fragments of isoform JM-A CYT-2 suggesting a prevalence of E4ICD2 over E4ICD1.
    Ligand-binding increases phosphorylation on tyrosine residues. Isoform JM-A CYT-2 is constitutively phosphorylated on tyrosine residues in a ligand-independent manner. E4ICD2 but not E4ICD1 is phosphorylated on tyrosine residues.
    Ubiquitinated. The ERBB4 intracellular domain is ubiquitinated and targeted to proteosomal degradation during mitosis mediated by the APC/C complex. Isoform JM-A CYT-1 and isoform JM-B CYT-1 are ubiquitinated by WWP1. The ERBB4 intracellular domain (E4ICD1) is ubiquitinated, and this involves NEDD4.
  • Cellular localization

    Membrane and Nucleus. Following proteolytical processing E4ICD (E4ICD1 or E4ICD2 generated from the respective isoforms) is translocated to the nucleus. Significantly more E4ICD2 than E4ICD1 is found in the nucleus. E4ICD2 colocalizes with YAP1 in the nucleus.
  • Target information above from: UniProt accession Q15303 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2066 Human
    • Omim: 600543 Human
    • SwissProt: Q15303 Human
    • Unigene: 390729 Human
    • Alternative names

      • 4ICD antibody
      • ALS19 antibody
      • Avian erythroblastic leukemia viral oncogene homolog 4 antibody
      • Avian erythroblastic leukemia viral v erb b2 oncogene homolog 4 antibody
      • E4ICD antibody
      • EC 2.7.10.1 antibody
      • ErbB 4 antibody
      • Erbb4 antibody
      • ERBB4 intracellular domain antibody
      • ERBB4_HUMAN antibody
      • HER 4 antibody
      • HER4 antibody
      • human epidermal growth factor receptor 4 antibody
      • Mer4 antibody
      • MGC138404 antibody
      • Oncogene ERBB4 antibody
      • p180erbB4 antibody
      • Proto-oncogene-like protein c-ErbB-4 antibody
      • Receptor protein tyrosine kinase erbB 4 precursor antibody
      • Receptor tyrosine protein kinase erbB 4 antibody
      • s80HER4 antibody
      • Tyrosine kinase type cell surface receptor HER4 antibody
      • Tyrosine kinase-type cell surface receptor HER4 antibody
      • v erb a avian erythroblastic leukemia viral oncogene homolog like 4 antibody
      • v erb a erythroblastic leukemia viral oncogene homolog 4 antibody
      • v-erb-a erythroblastic leukemia viral oncogene homolog 4 (avian) antibody
      • V-ERB-B2 avian erythroblastic leukemia viral oncogene homolog 4 antibody
      • Verba avian erythroblastic leukemia viral oncogene homolog like 4 antibody
      • Verba erythroblastic leukemia viral oncogene homolog 4 antibody
      • VERBB2 antibody
      see all

    Images

    • Western blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
      Western blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
      All lanes : Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273) at 1/1000 dilution

      Lane 1 : Untreated A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
      Lane 2 : A431 treated with 50mM pervanadate for 5 min whole cell lysate
      Lane 3 : A431 treated with 50mM pervanadate for 5 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour

      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: 147 kDa
      Observed band size: 180 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer: 5% NFDM/TBST

    • Western blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
      Western blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
      All lanes : Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273) at 1/1000 dilution

      Lane 1 : Untreated A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
      Lane 2 : A431 treated with 100 ng/ml Epidermal Growth Factor (EGF) for 30 min whole cell lysate
      Lane 3 : A431 treated with 100 ng/ml Epidermal Growth Factor (EGF) for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour

      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: 147 kDa
      Observed band size: 180 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer: 5% NFDM/TBST

    • Dot Blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
      Dot Blot - Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)

      Dot blot analysis using 1/1000 dilution ab109273 and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051)  secondary at 1/100000 dilution. 

      Blocking and diluting buffer: 5% NFDM/TBST

      Lane 1: ErbB4 non-phospho peptide

      Lane 2: ErbB4 Y1284 phospho peptide

      Lane 3: ErbB2 non-phospho peptide

      Lane 4: ErbB2 Y1248 phospho peptide

      Exposure time: 3 minutes

    • Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)
      Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)] (ab109273)

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (6)

    Publishing research using ab109273? Please let us know so that we can cite the reference in this datasheet.

    ab109273 has been referenced in 6 publications.

    • Yan T  et al. Multi-region sequencing unveils novel actionable targets and spatial heterogeneity in esophageal squamous cell carcinoma. Nat Commun 10:1670 (2019). PubMed: 30975989
    • Liu M  et al. Neuregulin-1 attenuates experimental cerebral malaria (ECM) pathogenesis by regulating ErbB4/AKT/STAT3 signaling. J Neuroinflammation 15:104 (2018). PubMed: 29636063
    • Chen J  et al. A role for ErbB signaling in the induction of reactive astrogliosis. Cell Discov 3:17044 (2017). PubMed: 29238610
    • Shekhar A  et al. Transcription factor ETV1 is essential for rapid conduction in the heart. J Clin Invest 126:4444-4459 (2016). IHC-Fr ; Mouse . PubMed: 27775552
    • Liang X  et al. Activation of NRG1-ERBB4 signaling potentiates mesenchymal stem cell-mediated myocardial repairs following myocardial infarction. Cell Death Dis 6:e1765 (2015). PubMed: 25996292
    • Slanina H  et al. Role of epidermal growth factor receptor signaling in the interaction of Neisseria meningitidis with endothelial cells. Infect Immun 82:1243-55 (2014). PubMed: 24379285

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293 cells transfected with ErbB2-EGFP)
    Gel Running Conditions
    Reduced Denaturing (Tricine-SDS-PAGE, 7% gel)
    Loading amount
    50 µg
    Treatment
    0.9% serum starved ~20 h, then stimulated with 20 ng/ml EGF for 5 min
    Specification
    293 cells transfected with ErbB2-EGFP
    Blocking step
    Milk as blocking agent for 50 minute(s) · Concentration: 5% · Temperature: ~22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Yan Cui

    Verified customer

    Submitted Dec 16 2020

    Question

    which ICC/IF protocol was used?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 18 2012

    Answer

    Thank you for contacting us.

    The lab has sent me the following protocol and information:

    The ICC protocol used to test this antibody is attached. Cells are serum-starved for overnight, and then treated with 100 ng/ml EGF for 5 min at 37 deg C.

    Immunofluorescent / Immunocytochemistry Protocol

    1. Solutions and reagents

    1.1. Washing buffer: PBST washing buffer: 1X PBS / 0.1% Tween-20 (Dulbecco’s Phosphate Buffered Saline)

    1.2. 2% Paraformaldehyde (prepare fresh by dissolving paraformaldehyde in 1X PBS by heating at 70ºC until dissolving. Use cold.)

    1.3. 0.1% Triton X-100

    1.4. Blocking buffer: PBS (Dulbecco’s Phosphate Buffered Saline) + 10% serum (serum origin depends on the host of the secondary antibody)

    1.5. Mounting medium for fluorescence.

    1.6 Acetone (Production Grade)

    2. Protocol

    2.1. Grow cells in chamber slides or in 6-well tissue plates containing sterile coverslips. Prior to fixation, cells should not reach more than 80% confluency.

    2.2. Fixation

    2.2.1. Remove medium from chamber slides or 6-well plates. Wash once with PBS-T.

    2.2.2. Fix cells with 2% paraformaldehyde for 20 min. For HeLa cells only, fix with acetone at -20 ºC for 5 mintues and continue to step 2.2.5.

    2.2.3. Wash cells twice with 1XPBST.

    2.2.4. Permeabilize cells with 0.1% Triton-X100 for 5 min

    2.2.5. Wash cells twice with 1XPBST.

    2.3. Blocking

    2.3.1. Block cells with blocking buffer for 1 hour at room temperature (option - O/N 4ºC).

    2.4. Staining

    2.4.1. Dilute primary antibody in the blocking buffer per recommendation on the data sheet.

    2.4.2. Apply primary antibody on the cells and incubate overnight at 4ºC.

    2.4.3. Wash cells twice with 1X PBST.

    2.4.4. Incubate cells in a dilution of fluorescently-labeled secondary antibody in PBS for 45 min at room temperature in the dark.

    2.4.5. Wash cells three times with 1X PBST.

    2.4.6. Counterstain cells with DAPI in a concentration of 300 nM in PBS for 5 min in the dark.

    2.4.7. Wash cells three times with 1X PBST.

    2.4.8. Mount slides with medium for fluorescent staining.

    2.4.9. Store slides in the dark.

    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

    Abcam Scientific Support

    Answered on Apr 18 2012

    Question

    Please suggest sopme ErbB4 antibodies that will detect the extracellular domain. For human samples and ICC, live and fixed cells.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 02 2011

    Answer

    Thank you for your telephone call earlier today. According to Swissprot/Uniprot protein database site, the extracellular domain of the ErbB 4 is from amino acids 26 – 651. http://www.uniprot.org/uniprot/Q15303 We do have the following ErbB4 antibodies tested in ICC-If and human, but regrettably, their immunogen seuqences are all in the cytoplasmic domain. They have been tested in ICC on fixed cells. As discussed on the telephone, I am sorry they will not have been tested and guarnateed for live cell imaging. I am copying the details below in case they are still of interest to you. I can recommend to review the online datasheets for further information. ab19391 Anti-ErbB 4 antibody [HFR1] Mouse monoclonal Applications: Flow Cyt, ICC/IF, IHC-P, IP, WB Reacts with: Human, Mouse Immunogen: RSTLQHPDYLQEYST amino acids 1250-1264 Click here (or use the following: https://www.abcam.com/ErbB4--HER4-antibody-HFR-1-ab19391.html). ab109273 Anti-ErbB 4 (phospho Y1284) antibody [EPR2273(2)] Rabbit monoclonal Applications: Flow Cyt, ICC/IF, WB Reacts with: Human, Mouse, Rat Immunogen: residues surrounding phosphorylated Tyrosine 1284 Click here (or use the following: https://www.abcam.com/ErbB4--HER4-phospho-Y1284-antibody-EPR22732-ab109273.html). ab61059 Anti-ErbB 4 (phospho Y1284) antibody Rabbit polyclonal Applications: ELISA, ICC/IF, WB Reacts with: Human, Mouse, Rat Immunogen: phosphorylation site of tyrosine 1284 (P-E-YP-L-S) Click here (or use the following: https://www.abcam.com/ErbB4--HER4-phospho-Y1284-antibody-ab61059.html). ab63354 Anti-ErbB 4 antibody Rabbit polyclonal Applications: ELISA, ICC/IF, IHC-P, WB Reacts with: Human Immunogen: phosphorylation site of tyrosine 1284. Click here (or use the following: https://www.abcam.com/ErbB4--HER4-antibody-ab63354.html). I am sorry we do not have antibodies that exactly meet your requirements on this occasion, but I hope this will still be helpful. If you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    Answered on Sep 02 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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