Rabbit Recombinant Monoclonal EGFR phospho T669 antibody. Carrier free. Suitable for Dot, WB, ICC/IF and reacts with Synthetic peptide, Human samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
Flow Cyt | Dot | WB | IHC-P | ICC/IF | |
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Human | Not recommended | Expected | Tested | Not recommended | Tested |
Synthetic peptide | Not recommended | Tested | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
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Species Human, Synthetic peptide | Dilution info - | Notes - |
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Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human, Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Synthetic peptide | Dilution info - | Notes - |
Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses (PubMed:10805725, PubMed:27153536, PubMed:2790960, PubMed:35538033). Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF (PubMed:12297049, PubMed:15611079, PubMed:17909029, PubMed:20837704, PubMed:27153536, PubMed:2790960, PubMed:7679104, PubMed:8144591, PubMed:9419975). 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 (PubMed:27153536). May also activate the NF-kappa-B signaling cascade (PubMed:11116146). 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 (PubMed:11602604). Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin (PubMed:11483589). Positively regulates cell migration via interaction with CCDC88A/GIV which retains EGFR at the cell membrane following ligand stimulation, promoting EGFR signaling which triggers cell migration (PubMed:20462955). Plays a role in enhancing learning and memory performance (By similarity). Plays a role in mammalian pain signaling (long-lasting hypersensitivity) (By similarity). Isoform 2 may act as an antagonist of EGF action. (Microbial infection) Acts as a receptor for hepatitis C virus (HCV) in hepatocytes and facilitates its cell entry. Mediates HCV entry by promoting the formation of the CD81-CLDN1 receptor complexes that are essential for HCV entry and by enhancing membrane fusion of cells expressing HCV envelope glycoproteins.
ERBB, ERBB1, HER1, EGFR, Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1
Rabbit Recombinant Monoclonal EGFR phospho T669 antibody. Carrier free. Suitable for Dot, WB, ICC/IF and reacts with Synthetic peptide, Human samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
Recognises EGFR phosphorylated on Threonine 669 of the mature human isoform 1 (corresponding to T693 from the precursor form P00533-1/p170)
ab284728 is the carrier-free version of Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980.
This product has switched from a hybridoma to recombinant production method on 9th February 2024.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
EGFR or Epidermal Growth Factor Receptor is a transmembrane glycoprotein that acts as a receptor for members of the epidermal growth factor family. Known alternatively as ErbB1 or HER1 this receptor has an approximate molecular weight of 170 kDa. EGFR is expressed in various cell types notably on epithelial cells and can influence multiple cellular processes through its kinase activity. It participates in the regulation of cell growth multiplication and survival by activating its kinase domain upon ligand binding.
The EGFR protein plays an important role in cellular communication and signaling processes. EGFR pairs with other receptor family members to form active dimers or even higher-order complexes which in turn initiate intracellular signaling cascades. Through these complexes EGFR influences many processes including cell differentiation and repair. This function of EGFR makes it an integral part of mammalian biology affecting how cells respond to their environment by mediating changes in gene expression.
EGFR is a central player in the MAPK and PI3K/Akt signaling pathways. Alongside other protein partners like KRAS and PI3 kinase it contributes to transmitting signals from the cell surface to the nucleus affecting gene transcription and cell behavior. These pathways are important for normal cell growth and division and aberrations in these pathways can lead to excessive or insufficient cell proliferation.
EGFR is pertinent to cancer biology including non-small cell lung cancer and glioblastoma where mutations or overexpression of the receptor frequently occur. It connects to proteins such as PTEN and BRAF which influence tumor progression and response to targeted therapies. EGFR's involvement in these disorders highlights its significance as a therapeutic target since it can be manipulated to alter disease progression.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This data was developed using Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980, the same antibody clone in a different buffer.
Dot blot analysis of EGFR (phospho T669) using Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/2500 dilution. Blocking and dilution buffer: 5% NFDM/TBST. Exposure time: 41s.
All lanes: Dot Blot - Anti-EGFR (phospho T669) antibody [EP2256Y] (Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980) at 1/1000 dilution
Lane 1: EGFR (phospho T669) peptide
Lane 2: EGFR non-phospho peptide
All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/2500 dilution
This data was developed using Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980, the same antibody clone in a different buffer.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-EGFR (phospho T669) antibody [EP2256Y] (Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980) at 1/2000 dilution
Lane 1: Untreated A431 (Human epidermoid carcinoma) whole cell lysate at 15 µg
Lane 2: A431 treated with 100ng/ml Epidermal Growth Factor (EGF) for 30mins whole cell lysate at 15 µg
Lane 3: A431 treated with 100ng/ml Epidermal Growth Factor (EGF) for 30mins whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 170 kDa
Exposure time: 40s
This data was developed using Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980, the same antibody clone in a different buffer.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 cells labelling EGFR with Anti-EGFR (phospho T669) antibody [EP2256Y] ab75980 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counterstain was DAPI (Blue).
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