Recombinant Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
- Datasheet
- References (27)
- Protocols
Overview
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Product name
Anti-EGFR (phospho Y1068) antibody [Y38]
See all EGFR primary antibodies -
Description
Rabbit monoclonal [Y38] to EGFR (phospho Y1068) -
Host species
Rabbit -
Specificity
ab32430 recognises only EGFR phosphorylated on Tyrosine 1068 of the mature human isoform 1 (corresponding to Y1092 from the precursor form P00533-1/p170). -
Tested applications
Suitable for: WB, ICC/IF, IPmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human, Pig -
Immunogen
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Positive control
- Purchase matching WB positive control:Recombinant human EGFR protein
- WB: A431 cell lysate treated with EGF. ICC/IF: A431 cell lysate treated with EGF.
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General notes
Mouse, 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.
A trial size is available to purchase for this antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y38 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab32430 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. Predicted molecular weight: 135 kDa. A blocking peptide is available for this antibody as ab205609. |
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| ICC/IF | 1/100 - 1/250. | |
| IP | 1/30. For unpurified format use at 1/150 dilution. |
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
- Entrez Gene: 397070 Pig
- 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 Y1068) antibody [Y38] (ab32430)](https://www.abcam.com/ps/products/32/ab32430/Images/ab32430-344788-Anti-egfr-phosphoY1068-antibody-Y38-westernblot-human.jpg)
Western blot - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)All lanes : Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430) at 1/1000 dilution (Purified)
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates
Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 minutes whole cell lysates
Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 minutes. Then the membrane was incubated with phosphatase
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: 135 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted? -
![Immunoprecipitation - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)](https://www.abcam.com/ps/products/32/ab32430/Images/ab32430-8-antiegfrphosphoY1068antibodyY38-immunoprecipitation-a431-human.jpg)
Immunoprecipitation - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)ab32430 (purified) at 1/30 dilution (2ug) immunoprecipitating EGFR in A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug . A431 (Human epidermoid carcinoma epithelial cell) treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
Lane 2 (+): ab32430 & A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32430 in A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
![Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)](https://www.abcam.com/ps/products/32/ab32430/Images/ab32430-255728-anti-egfr-phospho-y1068-antibody-y38-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)Immunocytochemistry/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling EGFR (phospho Y1068) with purified ab32430 (left) and EGFR with ab52894 (right) both at a dilution of 1/1000.
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
The image shows increased membrane staining after EGF (100ng/ml, 30min) treatment on A431 cells. The LP treatment decreased the increased membrane staining caused by EGF.
ab52894 was used as a Pan control for ab32430. The results showed membrane and cytoplasmic staining on untreated, EGF and EGF + LP treated A431 cells.
References
This product has been referenced in:
- Zhang Y et al. Inactivation of ADAMTS18 by aberrant promoter hypermethylation contribute to lung cancer progression. J Cell Physiol 234:6965-6975 (2019). Read more (PubMed: 30417422) »
- Ahmed M et al. Reading Conformational Changes in Proteins with a New Colloidal-Based Interfacial Biosensing System. ACS Appl Mater Interfaces 11:11125-11135 (2019). Read more (PubMed: 30799601) »
