Anti-EGFR antibody [EGFR1] (ab30)
- Datasheet
- References (16)
- Protocols
Overview
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Product nameAnti-EGFR antibody [EGFR1]
See all EGFR primary antibodies -
DescriptionMouse monoclonal [EGFR1] to EGFR
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Host speciesMouse
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Specificity
Recognises the external EGF-binding domain of the EGFR transmembrane glycoprotein. No effect on tyrosine kinase activity of EGFR. This antibody does not react with erbB-2, erbB-3 and erbB-4.
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Tested applicationsSuitable for: IHC-Fr, ICC/IF, IP, Flow Cyt, IHC - Wholemountmore details
Unsuitable for: ELISA or WB -
Species reactivityReacts with: Mouse, Horse, Human
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Immunogen
Tissue, cells or virus corresponding to Human EGFR (extracellular). Human epidermoid carcinoma line A431; epitope mapped between aa 6-273 of human EGFR.
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EpitopeExtracellular.
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Positive control
- IHC-Fr: Frozen normal human placenta ICC/IF: A431 cells Flow Cyt: A 431 cells
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General notes
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 orders@abcam.com.
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
PurityProtein G purified
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ClonalityMonoclonal
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Clone numberEGFR1
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MyelomaP3-NS1/1-Ag4-1
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IsotypeIgG2b
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Light chain typekappa
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Research areas
Associated products
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Affibody® Molecule
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-Fr | Use a concentration of 10 µg/ml. | |
| ICC/IF | Use a concentration of 5 µg/ml. | |
| IP | Use at an assay dependent concentration. | |
| Flow Cyt | Use a concentration of 1 µg/ml. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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| IHC - Wholemount | Use at an assay dependent concentration. |
Target
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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.
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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. -
Post-translational
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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Database links
- Entrez Gene: 1956 Human
- Entrez Gene: 13649 Mouse
- Omim: 131550 Human
- SwissProt: P00533 Human
- SwissProt: Q01279 Mouse
- Unigene: 488293 Human
- Unigene: 605083 Human
- Unigene: 420648 Mouse
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Alternative names
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
- Cell growth inhibiting protein 40 antibody
- Cell proliferation inducing protein 61 antibody
see all
Images
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![Flow Cytometry - Anti-EGFR antibody [EGFR1] (ab30)](https://www.abcam.com/ps/products/0/ab30/Images/ab30-336936-ab30FLOW1.jpg)
Flow Cytometry - Anti-EGFR antibody [EGFR1] (ab30)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.
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![Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EGFR1] (ab30)](https://www.abcam.com/ps/products/0/ab30/Images/ab30-336919-ICCAP332053101.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EGFR1] (ab30)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).
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![Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [EGFR1] (ab30)](https://www.abcam.com/ps/products/0/ab30/Images/ab30-331556-AP3320531hufrozenplacenta10ugml.jpg)
Immunohistochemistry (Frozen sections) - Anti-EGFR antibody [EGFR1] (ab30)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.
Protocols
Datasheets and documents
References
This product has been referenced in:
- 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 . Read more (PubMed: 29452913) »
- Minutti CM et al. Epidermal Growth Factor Receptor Expression Licenses Type-2 Helper T Cells to Function in a T Cell Receptor-Independent Fashion. Immunity 47:710-722.e6 (2017). Read more (PubMed: 29045902) »
