Anti-EGFR antibody (ab2430)
- Product nameAnti-EGFR antibodySee all EGFR primary antibodies ...
- DescriptionRabbit polyclonal to EGFR
- Tested applicationsFlow Cyt, ICC/IF, Sandwich ELISA, ICC, WB, IP, IHC-P, IHC-Fr, IHC-FrFl, In-Cell ELISA more details
- Species reactivityReacts with: Mouse, Rat, Sheep, Cow, Dog, Human
Synthetic peptide: EDMDDVVDADEY, corresponding to C terminal amino acids 1005-1016 of EGFR.
- Positive control
- Storage instructionsStore at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
- Storage bufferPreservative: 0.1% Sodium Azide
Constituents: 0.2% Gelatin, PBS
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
Our Abpromise guarantee covers the use of ab2430 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: Use at an assay dependent dilution.|
|ICC/IF||ICC/IF: 1/20 - 1/100. PubMed: 17567803|
|Sandwich ELISA||sELISA: Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [F4] to EGFR (ab62). For sandwich ELISA, use this antibody as Detection at 0.5µg/ml with Ab62 as Capture.|
|ICC||ICC: Use at an assay dependent dilution.|
|WB||WB: 1/200. Can be blocked with EGFR peptide (ab2432).|
|IP||IP: Use at an assay dependent dilution.|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|IHC-Fr||IHC-Fr: Use at an assay dependent dilution.|
|IHC-FrFl||IHC-FrFl: Use at an assay dependent concentration.|
|In-Cell ELISA||In-Cell ELISA: Use at an assay dependent concentration. PubMed: 21536028|
- 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 images
ab2430 at a 1/200 dilution staining A431 whole cell lysate.
Cells were stained with anti-EGFR (ab2430; red) and DAPI (blue). A 1/40 dilution of ab2430 was used.HeLa cells were fixed with 4% formaldehyde in PEM buffer. The coverslip was incubated in blocking buffer of 5% powdered milk in TBS-T plus 0.02% sodium azide for 1 hour at room temperature. Blocking buffer was removed and primary antibody was added at a dilution of 1/40 and incubated overnight at 4 degrees celsius. The coverslips were then washed 4-5 times with blocking buffer for 5 minutes. Secondary antibody was added at a dilution of 1/1000 and incubated at room temperature for one hour. From this point on coverslips were covered with foil to protect them from light. They were washed 5 times with TBS-T and then one time with PEM, for 5 minutes each wash. The coverslips were fixed 10-30 minutes in 4% formaldehyde in PEM buffer, then washed 3 times with PEM buffer for 5 minutes. 0.1M ammonium chloride in PEM buffer was added for 10 minutes to quench auto-florescence, and then slips were washed 2 times for 5 minutes in PEM followed by 3 washes for 5 minutes in TBS-T. Coverslips were then counterstained with DAPI in TBS-T for 1-2 minutes, TBS-T was then added and the coverslips mounted.
ab2430 staining formalin fixed paraffin-embedded sections of human adhesion tissue. The section was subjected to heat-mediated antigen retrieval in citrate buffer (pH 6.0) and blocked with 10% BSA for 1 hour prior to incubating with the primary antibody (diluted 1/200) for 12 hours at 4°C. A biotinylated goat anti-rabbit antibody was used as the secondary.
ab2430 staining EGFR in human stomach tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in 10 mM Citrate pH 6.0 and then blocking with 5% serum for 1 hour at 23°C was performed. The primary antibody was diluted 1/100 and incubated with sample for 1 hour at 23°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.
ab2430 immunoprecipitate of EGFR in human AGS whole cell lysate. 100 µg of cell lysate was incubated with primary antibody (1/100 in RIPA buffer) and matrix (Protein A/G) for 16 hours at 4°C.
For western blotting Ab6721 (1/1000) was used.
References for Anti-EGFR antibody (ab2430)
This product has been referenced in:
- 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 . Read more (PubMed: 23497505) »
- Tzouvelekis A et al. Increased expression of epidermal growth factor receptor (EGF-R) in patients with different forms of lung fibrosis. Biomed Res Int 2013:654354 (2013). IHC ; Human . Read more (PubMed: 23841084) »