Anti-EGFR (phospho Y1173) antibody [9H2] (ab24912)
- Product nameAnti-EGFR (phospho Y1173) antibody [9H2]See all EGFR primary antibodies ...
- DescriptionMouse monoclonal [9H2] to EGFR (phospho Y1173)
- SpecificityThis is a phosphorylation site-specific monoclonal antibody. It does not interact with the non-phosphorylated EGFR nor with unrelated Tyrosine-phosphorylated proteins.
- Tested applicationsWB, IP, ELISA, Flow Cyt, ICC, ICC/IF, IHC-Fr more details
- Species reactivityReacts with: Mouse, Rat, Dog, Human
Phosphopeptide conjugate (Human).
- EpitopeThis antibody specifically interacts with the 1170 - NAEpYLRV motif corresponding to the major autophosphorylation site of human EGFR.
- Storage instructionsStore at -20°C. Stable for 12 months at -20°C
- Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 2X [PBS, Polyethylene glycol, Sucrose]
- Concentration information loading...
- PurityPreadsorbed IgG fraction
- Purification notesThiophilic adsorption and size exclusion chromatography.
- Clonality Monoclonal
- Clone number9H2
- Light chain typekappa
Our Abpromise guarantee covers the use of ab24912 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||WB: Use a concentration of 0.5 µg/ml. Predicted molecular weight: 134 kDa.Can be blocked with EGFR (phospho Y1173) peptide (ab87004).|
|IP||IP: Use 1-10µg for 106 cells.|
|ELISA||ELISA: Use a concentration of 0.05 µg/ml.|
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
|ICC||ICC: Use at an assay dependent concentration.|
|ICC/IF||ICC/IF: Use a concentration of 0.1 - 1 µg/ml.|
|IHC-Fr||IHC-Fr: Use at an assay dependent concentration. PubMed: 20575069|
- 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 Y1173) antibody [9H2] images
Overlay histogram showing A431 cells stained with ab24912 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab24912, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
All lanes : Anti-EGFR (phospho Y1173) antibody [9H2] (ab24912) at 0.5 µg/ml
Lane 1 : Whole cell extracts of control (co) A549 tumor cells
Lane 2 : Whole cell extracts of EGF stimulated (EGF) A549 tumor cells
Lane 3 : Whole cell extracts of pervanadate treated (VH) A549 tumor cells
Lysates/proteins at 20000 cells per lane.
A HRP-conjugated antibody
developed using the ECL technique
Predicted band size : 134 kDa
Exposure time : 30 secondsRecommended blocking buffer CPPT: 0.5% (w/v) casein, 1% (w/v) PEG 4000, 1% (w/v) Polyvinylpyrrolidone (PVP), 0.1% Tween 20, 10mM Tris/HCl, pH 7.4, 150mM Sodium Chloride.
References for Anti-EGFR (phospho Y1173) antibody [9H2] (ab24912)
This product has been referenced in:
- Bradford D et al. Characterization of the netrin/RGMa receptor neogenin in neurogenic regions of the mouse and human adult forebrain. J Comp Neurol 518:3237-53 (2010). IHC-Fr ; Mouse . Read more (PubMed: 20575069) »
- Lee MS et al. ErbB-2 signaling is involved in regulating PSA secretion in androgen-independent human prostate cancer LNCaP C-81 cells. Oncogene 22:781-96 (2003). Read more (PubMed: 12569372) »