Anti-EGFR antibody [F4] (ab62)
- Product nameAnti-EGFR antibody [F4]See all EGFR primary antibodies ...
- DescriptionMouse monoclonal [F4] to EGFR
- SpecificityNo reaction with erbB-2, erbB-3 and erbB-4.
- Tested applicationsFlow Cyt, Sandwich ELISA, IP, ELISA, IHC-P, RIA, IHC-Fr more details
- Species reactivityReacts with: Human
Does not react withMouse, Rat
Synthetic peptide: DVVDADEYLIPQ from Human EGFR
- EpitopeThe epitope is aa 985-996 in the cytoplasmic domain of human EGFR. Reacts with both phosphorylated and unphosphorylated EGFR.
- Positive controlThis antibody gave a positive result in IHC in the following FFPE tissue: Human normal placenta.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPhosphate buffered saline
- Concentration information loading...
- PurityProtein A purified
- Clonality Monoclonal
- Clone numberF4
- Light chain typekappa
Our Abpromise guarantee covers the use of ab62 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Flow Cyt: 1/50.|
|Sandwich ELISA||sELISA: Use a concentration of 5 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to EGFR (ab2430). For sandwich ELISA, use this antibody as Capture at 5µg/ml with ab2430 as Detection.|
|IP||IP: Use at an assay dependent concentration.|
|ELISA||ELISA: Use at an assay dependent concentration.|
|IHC-P||IHC-P: Use at an assay dependent concentration.|
|RIA||RIA: Use at an assay dependent concentration.|
|IHC-Fr||IHC-Fr: 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 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 antibody [F4] images
Overlay histogram showing A431 cells stained with ab62 (red line). The cells were fixed with 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 (ab62, 1/50 dilution) 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, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
IHC image of EGFR staining in Human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab62, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
References for Anti-EGFR antibody [F4] (ab62)
This product has been referenced in:
- Jaganathan S et al. A functional nuclear epidermal growth factor receptor, SRC and stat3 heteromeric complex in pancreatic cancer cells. PLoS One 6:e19605 (2011). Human . Read more (PubMed: 21573184) »
- Jing MX et al. Estrogen receptor-alpha promoter methylation in sporadic basal-like breast cancer of Chinese women. Tumour Biol 32:713-9 (2011). IHC-P ; Human . Read more (PubMed: 21625942) »