Key features and details
- Mouse monoclonal [EM-12] to EGFR (phospho Y992)
- Suitable for: Flow Cyt, WB, IP
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-EGFR (phospho Y992) antibody [EM-12]
See all EGFR primary antibodies
DescriptionMouse monoclonal [EM-12] to EGFR (phospho Y992)
Specificityab81440 recognises EGFR phosphorylated on tyrosine 992.
Tested applicationsSuitable for: Flow Cyt, WB, IPmore details
Species reactivityReacts with: Human
Synthetic phospho-peptide covering sequence around tyrosine 992 of Human EGFR.
- EGF-stimulated A431. A431, CALU-3, Jurkat cell lines.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Concentration information loading...
PurityProtein A purified
Purification notesPurified from hybridoma culture supernatant. Purity: > 95% (by SDS-PAGE).
Our Abpromise guarantee covers the use of ab81440 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 170 kDa (predicted molecular weight: 134 kDa).|
|IP||Use at an assay dependent concentration.|
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 diseaseLung cancer
Inflammatory skin and bowel disease, neonatal, 2
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. Dephosphorylated by PTPN1 and PTPN2.
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 occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.
Cellular localizationSecreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF).
- Information by UniProt
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All lanes : Anti-EGFR (phospho Y992) antibody [EM-12] (ab81440) at 1 µg/ml
Lane 1 : EGF-treated A431 cell line
Lane 2 : EGF-treated CALU-3 cell line
Lane 3 : EGF-treated MCF-7 cell line
Lane 4 : EGF-treated Jurkat cell line
Lane 5 : EGF-treated Ramos cell line
Performed under reducing conditions.
Predicted band size: 134 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Immunoprecipitation of EGFR from EGF-treated A431 cells by ab81440. The precipitates were immunoblotted with ab81440 at 1µg/ml and goat anti-mouse-HRP.
Immunoprecipitation of EGFR from EGF-treated A431 cells by ab81440. The precipitates were immunoblotted with a commercial anti-EGFR polyclonal and goat anti-rabbit-HRP.
ab81440 has been referenced in 5 publications.
- Deng QF et al. Cyclooxygenase-2 mediates gefitinib resistance in non-small cell lung cancer through the EGFR/PI3K/AKT axis. J Cancer 11:3667-3674 (2020). PubMed: 32284763
- Pei YF et al. Silencing of LAMC2 Reverses Epithelial-Mesenchymal Transition and Inhibits Angiogenesis in Cholangiocarcinoma via Inactivation of the Epidermal Growth Factor Receptor Signaling Pathway. Am J Pathol 189:1637-1653 (2019). PubMed: 31345467
- Dittmann K et al. New roles for nuclear EGFR in regulating the stability and translation of mRNAs associated with VEGF signaling. PLoS One 12:e0189087 (2017). PubMed: 29253018
- Needham SR et al. EGFR oligomerization organizes kinase-active dimers into competent signalling platforms. Nat Commun 7:13307 (2016). PubMed: 27796308
- Harrison H et al. Oestrogen increases the activity of oestrogen receptor negative breast cancer stem cells through paracrine EGFR and Notch signalling. Breast Cancer Res 15:R21 (2013). WB ; Human . PubMed: 23497505