This antibody gave a positive signal in both EGF treated A431 and MDA MB 361 whole cell lysates.
This antibody gave a positive result when used in the following methanol fixed cell lines: MCF-7.
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 - 0.00025 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 200 kDa (predicted molecular weight: 148 kDa).
Binds and is activated by neuregulins and NTAK.
Epithelial tissues and brain.
Involvement in disease
Defects in ERBB3 are the cause of lethal congenital contracture syndrome type 2 (LCCS2) [MIM:607598]; also called Israeli Bedouin multiple contracture syndrome type A. LCCS2 is an autosomal recessive neurogenic form of a neonatally lethal arthrogryposis that is associated with atrophy of the anterior horn of the spinal cord. The LCCS2 syndrome is characterized by multiple joint contractures, anterior horn atrophy in the spinal cord, and a unique feature of a markedly distended urinary bladder. The phenotype suggests a spinal cord neuropathic etiology.
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily. Contains 1 protein kinase domain.
Overexpressed in a subset of human mammary tumors.
The cytoplasmic part of the receptor may interact with the SH2 or SH3 domains of many signal-transducing proteins.
Ligand-binding increases phosphorylation on tyrosine residues and promotes its association with the p85 subunit of phosphatidylinositol 3-kinase.
ab101407 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab101407 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-ErbB 3 (phospho Y1289) antibody (ab101407)
All lanes : Anti-ErbB 3 (phospho Y1289) antibody (ab101407) at 1 µg/ml
Lane 1 : EGF-Stimulated A431 Whole Cell Lysate Lane 2 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate Lane 3 : EGF-Stimulated A431 Whole Cell Lysate with Immunising peptide at 1 µg/ml Lane 4 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate with Immunising peptide at 1 µg/ml Lane 5 : EGF-Stimulated A431 Whole Cell Lysate with Control peptide at 1 µg/ml Lane 6 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate with Control peptide at 1 µg/ml
Lysates/proteins at 15 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
ErbB 3 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab101407 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
ab101407 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.