Recombinant Anti-EGFR (phospho T654) antibody [EP2255Y] (ab75986)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2255Y] to EGFR (phospho T654)
- Suitable for: WB, Dot blot
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-EGFR (phospho T654) antibody [EP2255Y]
See all EGFR primary antibodies -
Description
Rabbit monoclonal [EP2255Y] to EGFR (phospho T654) -
Host species
Rabbit -
Specificity
Recognises EGFR phosphorylated on Threonine 654 of the mature human isoform 1 (corresponding to T678 from the precursor form P00533-1/p170)
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Tested applications
Suitable for: WB, Dot blotmore details
Unsuitable for: Flow Cyt,IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30 minutes whole cell lysate.
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General notes
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2255Y -
Isotype
IgG -
Research areas
Associated products
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75986 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 |
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WB |
1/1000 - 1/50000. Predicted molecular weight: 134 kDa.
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Dot blot |
1/1000.
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Notes |
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WB
1/1000 - 1/50000. Predicted molecular weight: 134 kDa. |
Dot blot
1/1000. |
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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All lanes : Anti-EGFR (phospho T654) antibody [EP2255Y] (ab75986) at 1/1000 dilution (Purified)
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30 minutes whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30 minutes whole cell lysate, then the membrane treated with Lambda Phosphotase for 1 hour
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?Blocking buffer: 5% NFDM/TBST
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Primary antibody dilution: 1/1000
Secondary antibody: goat anti-rabbit IgG, (H+L), peroxidase conjugated
Secondary antibody dilution: 1/2500
Blocking & dilution buffer: 5% NFDM/TBST
Lane 1 sample: EGFR (pT654) phospho peptide
Lane 2 sample: EGFR non-phospho peptide
Exposure time: 3 minutes
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab75986 has not yet been referenced specifically in any publications.