Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)
- Datasheet
- References (9)
- Protocols
Overview
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Product nameAnti-EGFR (phospho Y1086) antibody [Y39]
See all EGFR primary antibodies -
DescriptionRabbit monoclonal [Y39] to EGFR (phospho Y1086)
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Host speciesRabbit
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SpecificityThis antibody only detects EGFR phosphorylated on Tyrosine 1086 of the mature human isoform 1.
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Tested applicationsSuitable for: WB, IP, Dot blot, ICC/IFmore details
Unsuitable for: Flow Cyt or IHC -
Species reactivityReacts with: Human
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Immunogen
Synthetic peptide within Human EGFR. The exact sequence is proprietary.
(Peptide available asab179559) -
Positive control
- A431 cells.
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General notes
A trial size is available to purchase for this antibody.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityWhole Cell Lysate
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ClonalityMonoclonal
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Clone numberY39
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IsotypeIgG
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Research areas
Associated products
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Isotype control
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab32086 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 |
|---|---|---|
| WB | 1/1000 - 1/10000. Detects a band of approximately 170 kDa (predicted molecular weight: 134 kDa). | |
| IP | 1/150. | |
| Dot blot | Use at an assay dependent concentration. | |
| ICC/IF | 1/100 - 1/250. |
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
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
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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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![Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-268154-ab32086WB.jpg)
Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/3000 dilution
Lane 1 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysates
Lane 2 : A431 (Human epidermoid carcinoma cell line) treated with 100 ng/ml EGF for 10 minutes whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 134 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-271444-ab32086-ICC1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR (phospho Y1086) with ab32086 at 1/100, 3 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, a AlexaFluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000, 2 μg/ml. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclear stain was DAPI (blue).
The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.
For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.
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![Dot blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-270050-ab32086-Dot-blot.jpg)
Dot blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)Dot blot analysis of EGFR (phospho Y1086) phospho peptide (Lane 1), EGFR non-phospho peptide (Lane 2), labeling EGFR (phospho Y1086) with ab32068 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 5 seconds.
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![Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086_2.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)Immunofluorescent analysis of EGFR (phospho Y1086) expression in A431 cell culture, using 1/100 ab32086. -
![Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086_1.jpg)
Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)All lanes : Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086) at 1/10000 dilution
Lane 1 : Untreated A431 cell lysate
Lane 2 : A431 cell lysate treated with EGF
Predicted band size: 134 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Datasheets and documents
References
This product has been referenced in:
- Hashikawa KI et al. Dysfunction of the circadian transcriptional factor CLOCK in mice resists chemical carcinogen-induced tumorigenesis. Sci Rep 7:9995 (2017). Read more (PubMed: 28855649) »
- Lantermann AB et al. Inhibition of Casein Kinase 1 Alpha Prevents Acquired Drug Resistance to Erlotinib in EGFR-Mutant Non-Small Cell Lung Cancer. Cancer Res 75:4937-48 (2015). Read more (PubMed: 26490646) »
