Phosphotyrosine ErbB4 / HER4 ELISA Kit (ab279792)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
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Product name
Phosphotyrosine ErbB4 / HER4 ELISA Kit
See all ErbB4 / HER4 kits -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
Phosphotyrosine ErbB4 / HER4 ELISA Kit (ab279792) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated ErbB4 / HER4 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-ErbB4 / HER4. An anti-ErbB4 / HER4 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated ErbB4 / HER4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed, and biotinylated anti-phosphotyrosine antibody is used to detect only tyrosine-phosphorylated protein. After washing away unbound antibody, Streptavidin-Conjugated HRP is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of phospho-ErbB4 / HER4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
For research use only - not intended for diagnostic use.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 20X Wash Buffer Concentrate 1 x 25ml 2X Cell lysate buffer 1 x 10ml 300X Streptavidin-Conjugated HRP 1 vial 5X Assay Diluent 1 x 15ml Biotinylated anti-phosphotyrosine antibody 2 vials Pan-ErbB4 / HER4 Coated Microplate 1 unit Positive Control - T47D cell lysate 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
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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Alternative names
- 4ICD
- ALS19
- Avian erythroblastic leukemia viral oncogene homolog 4
see all -
Database links
- Entrez Gene: 2066 Human
- Omim: 600543 Human
- SwissProt: Q15303 Human
- Unigene: 390729 Human
Images
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Positive Control
T47D cells were treated with Pervanadate at 37°C for 10 mins.
Solubilize cells at 4 x 107 cells/ml in lysis buffer.
Serial dilutions of lysates were analyzed in this ELISA. .
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T47D cells treated/untreated with Pervanadate.T47D cells were treated or untreated with Pervanadate for 10 min at 37degC.