Recombinant Anti-ErbB4 / HER4 (phospho Y1162) antibody [EP2270Y] - BSA and Azide free (ab247386)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2270Y] to ErbB4 / HER4 (phospho Y1162) - BSA and Azide free
- Suitable for: WB, IHC-P, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ErbB4 / HER4 (phospho Y1162) antibody [EP2270Y] - BSA and Azide free
See all ErbB4 / HER4 primary antibodies -
Description
Rabbit monoclonal [EP2270Y] to ErbB4 / HER4 (phospho Y1162) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A431 treated with 100 ng/ml EGF. IHC-P: Human breast carcinoma tissue. IP: A431 cells.
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General notes
ab247386 is the carrier-free version of ab68478.
This product has switched from a hybridoma to recombinant production method on 31st May 2023.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2270Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247386 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 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 200 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 200 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
Target
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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
modificationsIsoform 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. - Information by UniProt
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Database links
- Entrez Gene: 2066 Human
- Omim: 600543 Human
- SwissProt: Q15303 Human
- Unigene: 390729 Human
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Alternative names
- 4ICD antibody
- ALS19 antibody
- Avian erythroblastic leukemia viral oncogene homolog 4 antibody
see all
Images
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All lanes : Anti-ErbB4 / HER4 (phospho Y1162) antibody [EP2270Y] (ab68478) at 1/5000 dilution
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 min whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 min whole cell lysate, then membrane treated with Alkaline 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: 200 kDa
Observed band size: 200 kDa
Exposure time: 10 secondsThis data was developed using ab68478 the same antibody clone in a different buffer.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
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This data was developed using ab68478 the same antibody clone in a different buffer.
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling ErbB4 / HER4 with ab68478 at 1/1000 dilution, followed by ab209101 Rabbit specific IHC polymer detection kit HRP/DAB. Positive staining on Human breast carcinoma without alkaline phosphatase treatment (A) compared to no signal detected when treated with alkaline phosphatase (B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval Solution2) for 30 mins. Counterstained with hematoxylin.
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This data was developed using ab68478 the same antibody clone in a different buffer.
ErbB4 / HER4 (phospho Y1162) was immunoprecipitated from 0.35 mg of A431 treated with EGF (100ng/ml 30min) whole cell lysate with ab68478 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab68478 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
Lane 1: A431 treated with EGF (100ng/ml 30min) whole cell lysate.
Lane 2: ab68478 IP in A431 treated with EGF (100ng/ml 30min) whole cell lysate.
Lane 3: Isotype control instead of ab68478 in A431 treated with EGF (100ng/ml 30min) whole cell lysate.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab247386 has not yet been referenced specifically in any publications.