Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2259Y] to EGFR (phospho S1046 + S1047) - BSA and Azide free
- Suitable for: WB, IP, ICC/IF, Dot blot
- Reacts with: Human
Product nameAnti-EGFR (phospho S1046 + S1047) antibody [EP2259Y] - BSA and Azide free
See all EGFR primary antibodies
DescriptionRabbit monoclonal [EP2259Y] to EGFR (phospho S1046 + S1047) - BSA and Azide free
Tested applicationsSuitable for: WB, IP, ICC/IF, Dot blotmore details
Unsuitable for: Flow Cyt or IHC-P
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human EGFR (phospho S1046 + S1047).
- WB: A431 cell lysates treated with EGF. ICC/IF: A431 cells treated with EGF.
ab256225 is the carrier-free version of ab76300 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
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.
Ab256225 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
Storage bufferpH: 7.2
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab256225 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 134 kDa).|
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|Dot blot||Use at an assay dependent concentration.|
FunctionReceptor 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 specificityUbiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
Involvement in diseaseLung cancer
Inflammatory skin and bowel disease, neonatal, 2
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain.
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 localizationSecreted 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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Immunocytochemistry/Immunofluorescence analysis of untreated and EGF (100ng/mL for 10 minutes) treated A431 cells labelling EGFR (phospho S1046 + S1047) with ab76300 at 1/100. 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. The cells were co-stained with ab195889, an Alexa Fluor® 594-conjugated mouse anti-tubulin (1/200). Nuclei counterstained with DAPI (blue).
Control 1: untreated A431 cells incubated with PBS instead of primary antibody followed by incubation with ab150077 Alexa Fluor® 488 goat anti-rabbit IgG.
Control 2: EGF treated A431 cells incubated with PBS instead of primary antibody followed by incubation with ab150077 Alexa Fluor® 488 goat anti-rabbit IgG.
This data was developed using the same antibody clone in a different buffer formulation containing Tris Glycine, BSA, glycerol, and sodium azide (ab76300).
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.
ab256225 has not yet been referenced specifically in any publications.