Key features and details
- Rabbit polyclonal to EGFR (phospho Y1148)
- Suitable for: WB, ICC
- Reacts with: Human
- Isotype: IgG
Product nameAnti-EGFR (phospho Y1148) antibody
See all EGFR primary antibodies
DescriptionRabbit polyclonal to EGFR (phospho Y1148)
Tested applicationsSuitable for: WB, ICCmore details
Species reactivityReacts with: Human
Synthetic phosphopeptide (Human) derived from the region of EGFR that contains tyrosine 1148.
- A431 cells stimulated with EGF, A549 cells.
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
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 1148.
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5651 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 140 kDa.
1/1000. Detects a band of approximately 140 kDa.
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 Y1148) antibody (ab5651) at 1/1000 dilution
Lane 1 : A431 cells unstimulated with no peptide
Lane 2 : A431 cells stimulated with 200 ng/mL EGF for 15 minutes with no peptide
Lane 3 : A431 cells stimulated with 200 ng/mL EGF for 15 minutes with non-phosphopeptide corresponding to the phosphopeptide immunogen
Lane 4 : A431 cells stimulated with 200 ng/mL EGF for 15 minutes with a generic phosphotyrosine-containing peptide
Lane 5 : A431 cells stimulated with 200 ng/mL EGF for 15 minutes with phosphopeptide immunogen
All lanes : goat F(ab’)2 anti-rabbit IgG HRP conjugate
Blocking buffer: 5% BSA-TBST
The phosphopeptide corresponding to EGFR [pY1148] blocks the signal.
Immunocytochemical analysis of EGFR (pY1148) on A549 cells. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.25% Triton™ X-100. The cells were labeled with ab5651 at 1/250 dilution in 1% BSA and incubated for 3 hours at room temperature followed by Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody at a dilution of 1/400 (Panel a: green). Nuclei (Panel b: blue) were stained DAPI. (Panel c: red) F-actin was stained with Alexa Fluor 594 Phalloidin. Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control.
ab5651 has been referenced in 1 publication.
- Liao HW et al. PRMT1-mediated methylation of the EGF receptor regulates signaling and cetuximab response. J Clin Invest 125:4529-43 (2015). PubMed: 26571401