Anti-EGFR (phospho Y845) antibody (ab5636)
- Product nameAnti-EGFR (phospho Y845) antibodySee all EGFR primary antibodies ...
- DescriptionRabbit polyclonal to EGFR (phospho Y845)
- Tested applicationsICC/IF, WB, ICC more details
- Species reactivityReacts with: Mouse, Rat, Human
Synthetic phosphopeptide derived from the region of EGFR that contains tyrosine 845.
- Positive control
- Human Epidermoid Carcinoma (A431) cells and cortic smooth muscle cells.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.05% Sodium Azide
Constituents: PBS, 1mg/ml BSA. pH 7.3
- Concentration information loading...
- PurityImmunogen affinity purified
- Purification notesThe antibody has been negatively preadsorbed using (i) a non phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated epidermal growth factor receptor (EGFR), and (ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine, irrespective of the sequence. The final product is generated by affinity chromatography using an EGFR-derived peptide that is phosphorylated at tyrosine 845.
- Clonality Polyclonal
Our Abpromise guarantee covers the use of ab5636 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: 1/50 (PMID 19147829).
WB: Use at a concentration of 0.1 - 1.0 µg/ml. Detects a band of approximately 185 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
- FunctionReceptor 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 specificityUbiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
- Involvement in diseaseDefects in EGFR are associated with lung cancer (LNCR) [MIM:211980]. LNCR is a common malignancy affecting tissues of the lung. The most common form of lung cancer is non-small cell lung cancer (NSCLC) that can be divided into 3 major histologic subtypes: squamous cell carcinoma, adenocarcinoma, and large cell lung cancer. NSCLC is often diagnosed at an advanced stage and has a poor prognosis.
- Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain.
modificationsPhosphorylation 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.
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 occur. Deubiquitinated by OTUD7B, preventing degradation.
Methylated. Methylation at Arg-1199 by PRMT5 positively stimulates phosphorylation at Tyr-1197.
- Cellular localizationSecreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand.
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Anti-EGFR (phospho Y845) antibody images
Peptide Competition: Extracts prepared from A431 cells stimulated with EGF were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA TBST buffer overnight at 4°C, then incubated with 0.40 µg/mL ab5636 antibody, for two hours at room temperature in a 3% BSA-TBST buffer, following its prior incubation with: no peptide (1), the phosphopeptide immunogen (2), the non-phosphopeptide corresponding to the immunogen (3), or a generic phosphotyrosine containing peptide (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the phosphopeptide corresponding to ab5636 blocks the antibody signal, demonstrating the specificity of the antibody.
Immunodetection using ab5636 in cultured cortic smooth muscle cells treated with Angiotensin II.
References for Anti-EGFR (phospho Y845) antibody (ab5636)
This product has been referenced in:
- Jaldety Y et al. Sperm EGFR mediates a7 AChR activation to promote fertilization. J Biol Chem : (2012). Read more (PubMed: 22577141) »
- Pontillo CA et al. Activation of c-Src/HER1/STAT5b and HER1/ERK1/2 signaling pathways and cell migration by hexachlorobenzene in MDA-MB-231 human breast cancer cell line. Toxicol Sci 120:284-96 (2011). Read more (PubMed: 21205633) »