Anti-EGFR (phospho Y992) antibody (ab5638)

Overview

• Product nameAnti-EGFR (phospho Y992) antibodySee all EGFR primary antibodies ...
• Description
  Rabbit polyclonal to EGFR (phospho Y992)
• Tested applicationsWB more details
• Species reactivity
  Reacts with: Human
  
• Immunogen

  Synthetic peptide phosphopeptide derived from the region of EGFR that contains tyrosine 992.

• Positive controlA431 cells stimulated with EGF; NIH 3T3 cells expressing human EGFR and stimulated with EGF.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPreservative: 0.05% Sodium Azide
  Constituents: 50% Glycerol, Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), 1mg/ml BSA(IgG, protease free) as a carrier. 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 922.
• Clonality Polyclonal
• IsotypeIgG
• Research Areas
  • Cancer
  • Tumor biomarkers
  • Receptors

  • Cancer
  • Oncoproteins/suppressors
  • Oncoproteins
  • Growth factor receptors

  • Cancer
  • Signal transduction
  • Protein phosphorylation
  • Tyrosine kinases
  • Receptor tyrosine kinases

  • Signal Transduction
  • Protein Phosphorylation
  • Tyrosine Kinases
  • Receptor Tyrosine Kinases

  • Signal Transduction
  • Growth Factors/Hormones
  • EGF

Applications

Target

• 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.
• Post-translational
  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.

Target information above from: UniProt accession P00533 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 1956 Human
  • Omim: 131550 Human
  • SwissProt: P00533 Human
  • Unigene: 488293 Human

• Alternative names
  Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibodyAvian erythroblastic leukemia viral (verbb) oncogene homolog antibodyCell growth inhibiting protein 40 antibody

  Cell proliferation inducing protein 61 antibodyEGF R antibodyEGFR antibodyEGFR antibodyEGFR_HUMAN antibodyEpidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) antibodyEpidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) antibodyEpidermal growth factor receptor antibodyerbb 1 antibodyErbb antibodyErbb1 antibodyERBB1 antibodyErrp antibodyHER1 antibodymENA antibodyOncogene ERBB antibodyPIG61 antibodyProto-oncogene c-ErbB-1 antibodyReceptor tyrosine protein kinase ErbB 1 antibodyReceptor tyrosine protein kinase ErbB1 antibodyReceptor tyrosine-protein kinase ErbB-1 antibodyUrogastrone antibodywa2 antibodyWa5 antibody

  see all

Anti-EGFR (phospho Y992) antibody images

• 

  Western blot - EGFR (phospho Y992) antibody (ab5638)
  Peptide Competition: Cell extracts prepared from NIH 3T3 cells expressing EGFR were starved for 30 hours, then stimulated for 10 minutes with 30 ng/mL EGF (+), or left unstimulated (-), then resolved by SDS-PAGE on a 6% Tris-glycine gel, and transferred to nitrocellulose. The membranes were then incubated with 0.50 µg/mL ab5638 antibody, following prior incubation in the absence of the peptide immunogen (1, 2), presence of the peptide immunogen (3, 4), or the non-phosphopeptide corresponding to the EGFR phosphopeptide (5, 6). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, demonstrating the specificity of the ab5638 antibody for this phosphorylated residue. The data also show the activation of the EGFR after stimulation with EGF.

Protocols

• Peptide Competition Experiment
• Western blotting protocol