Rabbit Recombinant Multiclonal EGFR phospho Y845 + Y1068 + Y1086 antibody. Carrier free. Suitable for Dot, ICC/IF, IP, WB and reacts with Synthetic peptide - Human, Human samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
Dot | ICC/IF | IP | WB | IHC-P | Flow Cyt | |
---|---|---|---|---|---|---|
Human | Expected | Tested | Tested | Tested | Not recommended | Not recommended |
Mouse | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Synthetic peptide - Human | Tested | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
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Species Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | 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 - Human, Mouse, Rat | 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 - Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Synthetic peptide - Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat, Human, Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human, Synthetic peptide - Human, Mouse, Rat | 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, Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1
Rabbit Recombinant Multiclonal EGFR phospho Y845 + Y1068 + Y1086 antibody. Carrier free. Suitable for Dot, ICC/IF, IP, WB and reacts with Synthetic peptide - Human, Human samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
ab319114 is the carrier-free version of Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113.
What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:
View our range of recombinant multiclonal antibodies.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.
Compatibility
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.
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.
EGFR (phospho Y845 + Y1068 + Y1086) Western blot staining using rabbit Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody
This data was developed using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
In Western blot, Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894) staining at 1/1000 dilution.
All lanes: Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113) at 1/5000 dilution
Lane 1: Untreated A431 (human epidermoid carcinoma epithelial cell) whole cell lysate (untreated membrane) at 10 µg
Lane 2: A431 treated with 100ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 10 µg
Lane 3: Untreated A431 whole cell lysate (alkaline phosphatase treated membrane) at 10 µg
Lane 4: A431 treated with 100ng/ml EGF for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 175 kDa, 124 kDa
Exposure time: 169s
EGFR (phospho Y845 + Y1068 + Y1086) Western blot staining using rabbit Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody
This data was developed using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 was shown to bind specifically to EGFR. Target of interest was observed at 175kDa, in wild-type HeLa cell lysates (lane 1/2/5/6) with no signal observed at this size in EGFR knockout cell line (lane 3/4/7/8) (knockout cell line Human EGFR knockout HeLa cell line ab255385 / knockout cell lysate Human EGFR knockout HeLa cell lysate ab263845).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894) staining at 1/1000 dilution.
All lanes: Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113) at 1/1000 dilution
Lane 1: Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 2: Wild-type HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 20 µg
Lane 3: Untreated EGFR knockout HeLa whole cell lysate (untreated membrane) at 20 µg
Lane 4: EGFR knockout HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 20 µg
Lane 5: Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 6: Wild-type HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 7: Untreated EGFR knockout HeLa whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 8: EGFR knockout HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under reducing conditions.
Observed band size: 175 kDa, 36 kDa
Exposure time: 180s
This data was developed using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113, the same antibody clone in a different buffer formulation.
Dot blot analysis of EGFR (phospho Y845 + Y1068 + Y1086) using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 at 1:1000 (0.496 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.
Lane1: Human EGFR (phospho Y1068) peptide
Lane2: Human EGFR non-phospho peptide
Lane3: Human EGFR (phospho Y1086) peptide
Lane4: Human EGFR non-phospho peptide
Lane5: Human EGFR (phospho Y845) peptide
Lane6: Human EGFR non-phospho peptide
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Dot Blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113) at 1/1000 dilution
Lane 1: Human EGFR (phospho Y1068) peptide
Lanes 2, 4 and 6: Human EGFR non-phospho peptide
Lane 3: Human EGFR (phospho Y1086) peptide
Lane 5: Human EGFR (phospho Y845) peptide
All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 180s
This data was developed using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling EGFR (phospho Y845 + Y1068 + Y1086) with Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 at 1/10000 (0.05 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing increased membranous staining in A431 cells treated with EGF (100 ng/ml) for 30 min (shown in green). The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
This data was developed using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113, the same antibody clone in a different buffer formulation.
EGFR (phospho Y845 + Y1068 + Y1086) was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate with Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate 2 ug
Lane 2: Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 IP in A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113 in A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] ab319113) at 1/30 dilution
All lanes: A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 15s
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