Key features and details
- Mouse monoclonal [EGFR1] to EGFR
- Suitable for: IHC-Fr, ICC/IF, Flow Cyt
- Reacts with: Horse, Human
- Isotype: IgG2b
参阅全部 EGFR 一抗
描述小鼠单克隆抗体[EGFR1] to EGFR
经测试应用适用于: IHC-Fr, ICC/IF, Flow Cytmore details
不适用于: ELISA or WB
种属反应性与反应: Horse, Human
Tissue, cells or virus corresponding to Human EGFR (extracellular). Human epidermoid carcinoma line A431; epitope mapped between aa 6-273 of human EGFR.
- IHC-Fr: Frozen normal human placenta ICC/IF: A431 cells Flow Cyt: A 431 cells
Recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR.
This antibody clone is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact email@example.com.
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
存储溶液Preservative: 0.02% Sodium azide
Concentration information loading...
纯度Protein G purified
Our Abpromise guarantee covers the use of ab30 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use a concentration of 10 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use a concentration of 1 µg/ml.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
功能Receptor 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.
组织特异性Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
Inflammatory skin and bowel disease, neonatal, 2
序列相似性Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain.
翻译后修饰Phosphorylation 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.
细胞定位Secreted 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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Overlay histograms showing left positive A431 cells and right negative Jurkat cells stained with ab30 (red line). The cells were incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab30, 1x106 cells in 100ul at 1ug/ml) for 30 min on ice. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) preabsorbed (ab150117) at 1/2000 dilution for 30 min on ice.
Isotype control antibody (black line) was mouse IgG2bκ (ab170192) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
ab30 staining EGFR (colored green) in A431 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab30 at 5μg/ml overnight at 4ºC. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
IHC image of EGFR staining in a section of frozen normal human placenta, fixed in 10% paraformaldehyde (10 min). Staining was 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 20mins. The section was then incubated with ab30, 10ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
ab30 被引用在 23 文献中.
- Lima Moura S et al. Matrix Effect in the Isolation of Breast Cancer-Derived Nanovesicles by Immunomagnetic Separation and Electrochemical Immunosensing-A Comparative Study. Sensors (Basel) 20:N/A (2020). PubMed: 32054015
- Liu C et al. Enhanced blood-brain-barrier penetrability and tumor-targeting efficiency by peptide-functionalized poly(amidoamine) dendrimer for the therapy of gliomas. Nanotheranostics 3:311-330 (2019). PubMed: 31687320
- Majorini MT et al. cIAP1 regulates the EGFR/Snai2 axis in triple-negative breast cancer cells. Cell Death Differ 25:2147-2164 (2018). PubMed: 29674627
- Fisher SA et al. Photo-immobilized EGF chemical gradients differentially impact breast cancer cell invasion and drug response in defined 3D hydrogels. Biomaterials 178:751-766 (2018). Flow Cyt ; Human . PubMed: 29452913
- Bäck N et al. The endocytic pathways of a secretory granule membrane protein in HEK293 cells: PAM and EGF traverse a dynamic multivesicular body network together. Eur J Cell Biol 96:407-417 (2017). PubMed: 28377049