Recombinant Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)
![Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-268154-anti-egfr-phospho-y1086-antibody-y39-western-blot.jpg "Western blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y39] to EGFR (phospho Y1086)
- Suitable for: WB, IP, Dot blot, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-EGFR (phospho Y1086) antibody [Y39]
See all EGFR primary antibodies -
Description
Rabbit monoclonal [Y39] to EGFR (phospho Y1086) -
Host species
Rabbit -
Specificity
This antibody only detects EGFR phosphorylated on Tyrosine 1086 of the mature human isoform 1. -
Tested applications
Suitable for: WB, IP, Dot blot, ICC/IFmore details
Unsuitable for: Flow Cyt or IHC -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human EGFR. The exact sequence is proprietary.
(Peptide available asab179559) -
Positive control
- A431 cells. IP: A431 cell lysate
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General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y39 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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).
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| IP |
1/150.
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| Dot blot |
Use at an assay dependent concentration.
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| ICC/IF |
1/100 - 1/250.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Detects a band of approximately 170 kDa (predicted molecular weight: 134 kDa). |
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IP
1/150. |
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Dot blot
Use at an assay dependent concentration. |
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ICC/IF
1/100 - 1/250. |
Target
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Function
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. -
Tissue specificity
Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers. -
Involvement in disease
Lung cancer
Inflammatory skin and bowel disease, neonatal, 2 -
Sequence similarities
Belongs 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 localization
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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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)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-anti-egfr-phospho-y1086-antibody-y39-immunofluorescence.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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![Immunoprecipitation - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-8-anti-egfr-phospho-y1086-antibody-y39-immunoprecipitation-a431-human.jpg)
Immunoprecipitation - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)Purified ab32086 at 1/20 dilution (1µg) immunoprecipitating EGFR in A431 treated with 100ng/mL EGF for 30min whole cell lysate.
Lane 1 (input): A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/mL EGF for 30min whole cell lysate 10µg
Lane 2 (+): ab32086 + A431 treated with 100ng/mL EGF for 30min whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32086 in A431 treated with 100ng/mL EGF for 30min whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 175 kDa -
![Dot Blot - Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-270050-anti-egfr-phospho-y1086-antibody-y39-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/ab32086-43840-anti-egfr-phospho-y1086-antibody-y39-immunofluorescence.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/ab32086-43841-anti-egfr-phospho-y1086-antibody-y39-western-blot.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? -
![Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)](https://www.abcam.com/ps/products/32/ab32086/Images/ab32086-7-benefits-of-recombinant-antibodies.png)
Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (10)
ab32086 has been referenced in 10 publications.
- Kenny FN et al. Tissue stiffening promotes keratinocyte proliferation through activation of epidermal growth factor signaling. J Cell Sci 131:N/A (2018). PubMed: 29669739
- Hashikawa KI et al. Dysfunction of the circadian transcriptional factor CLOCK in mice resists chemical carcinogen-induced tumorigenesis. Sci Rep 7:9995 (2017). 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). PubMed: 26490646
- Meneses-Lorente G et al. Preclinical pharmacokinetics, pharmacodynamics, and efficacy of RG7116: a novel humanized, glycoengineered anti-HER3 antibody. Cancer Chemother Pharmacol 75:837-50 (2015). WB . PubMed: 25702049
- Johansson M et al. The soluble form of the tumor suppressor Lrig1 potently inhibits in vivo glioma growth irrespective of EGF receptor status. Neuro Oncol 15:1200-11 (2013). WB ; Human . PubMed: 23723255
- Xu ZH et al. RAF1-MEK1-ERK/AKT axis may confer NSCLC cell lines resistance to erlotinib. Int J Clin Exp Pathol 6:1493-504 (2013). WB ; Human . PubMed: 23923067
- Bill A et al. Anti-proliferative effect of cytohesin inhibition in gefitinib-resistant lung cancer cells. PLoS One 7:e41179 (2012). WB . PubMed: 22815959
- Castoldi R et al. Molecular characterization of novel trispecific ErbB-cMet-IGF1R antibodies and their antigen-binding properties. Protein Eng Des Sel 25:551-60 (2012). WB ; Human . PubMed: 22936109
- Dai G et al. Maternal hepatic growth response to pregnancy in the mouse. Exp Biol Med (Maywood) 236:1322-32 (2011). PubMed: 21969712
- Kramer EL et al. Perinatal increases in TGF-{alpha} disrupt the saccular phase of lung morphogenesis and cause remodeling: microarray analysis. Am J Physiol Lung Cell Mol Physiol 293:L314-27 (2007). PubMed: 17468132
