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48-602MAG
Buffer Detection Kit for Magnetic Beads
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The epidermal growth factor receptor (EGFR; Proto-oncogene c-ErbB-1) is a ubiquitously receptor tyrosine kinase that is activated by a variety of ligands including EGF, EPGN, TGFA, and EREG. Ligand-bound EGFR undergoes dimerization and autophosphorylation exposing cytoplasmic domains which allow interaction with adaptor proteins and induction of several signaling pathways including PI3-AKT; PLCγ-PKC; Notch1; and Ras-Raf. EGFR therefore contributes to a diverse range of cellular processes inlcuding cell proliferation. Several studies have reported that EGFR signaling plays an important role in the development of various tumors. EGFR is regulated by PTPRJ and PTPRK phosphatases, and by the endocytic machinery including EPS15 that mediates internalization and degradation of the receptor.
References
Product Information
Format
Purified
Control
A431 cell lysate
Presentation
Purified mouse monoclonal IgG1κ in buffer containing PBS without preservatives.
Anti-EGFR Antibody, clone 225 (Azide-free) is an antibody against EGFR for use in Immunoprecipitation, Neutralizing.
Key Applications
Immunoprecipitation
Neutralizing
Application Notes
Neutralizing Assay Analysis: A representative lot was used by an independent laboratory in A431 cells. (Kawamoto, T., et al. (1984). Journal of Biological Chemistry. 259(12):7761-7766.)
Biological Information
Immunogen
Partially purified human EGFR from A431 cells
Clone
225
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
The protein encoded by this gene is a transmembrane glycoprotein that is a member of the protein kinase superfamily. This protein is a receptor for members of the epidermal growth factor family. EGFR is a cell surface protein that binds to epidermal growth factor. Binding of the protein to a ligand induces receptor dimerization and tyrosine autophosphorylation and leads to cell proliferation. Mutations in this gene are associated with lung cancer. Multiple alternatively spliced transcript variants that encode different protein isoforms have been found for this gene. [provided by RefSeq, Jul 2010].
FUNCTION: Isoform 2/truncated isoform may act as an antagonist.
SIZE: 1210 amino acids; 134277 Da
SUBUNIT: Binds RIPK1. CBL interacts with the autophosphorylated C- terminal tail of the EGF receptor. Part of a complex with ERBB2 and either PIK3C2A or PIK3C2B. The autophosphorylated form interacts with PIK3C2B, maybe indirectly. Interacts with PELP1.
SUBCELLULAR LOCATION: Cell membrane; Single-pass type I membrane protein. & Isoform 2: Secreted.
TISSUE SPECIFICITY: Expressed in placenta. Isoform 2 is also expressed in ovarian cancers.
PTM: Phosphorylation of Ser-695 is partial and occurs only if Thr- 693 is phosphorylated. & 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 occur.DISEASE: Defects in EGFR are associated with lung cancer [MIM:211980].
SIMILARITY: Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily. & Contains 1 protein kinase domain.
MISCELLANEOUS: Binding of EGF to the receptor leads to dimerization, internalization of the EGF-receptor complex, induction of the tyrosine kinase activity, stimulation of cell DNA synthesis, and cell proliferation.
Molecular Weight
~170 kDa observed. Uniprot gives a calculated molecular weight of 135 but can be observed at ~170 kDa due to phosphorylation. (Konishi, A., et al. (2003). The Journal of Biological Chemistry. 278(37):35049–35056.)
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Immunoprecipitation in A431 cell lysate.
Immunoprecipitation Analysis: 10 µg of this antibody immunoprecipitated EGFR from A431 cell lysate.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Relation of epidermal growth factor receptor concentration to growth of human epidermoid carcinoma A431 cells. Kawamoto, T, et al. J. Biol. Chem., 259: 7761-6 (1984)
1983
The relation between the concentration of epidermal growth factor (EGF) receptor/kinase and effects of EGF on cell proliferation has been studied using variant A431 cells and antagonist anti-EGF receptor monoclonal antibodies. Clonal A431 cell variants selected for escape from the EGF-mediated growth inhibition of parental A431 cells all have reduced concentrations of EGF receptor/kinase; Harvey sarcoma virus-transformed A431 cells, which have escaped from EGF-mediated growth inhibition, also have reduced EGF receptors. Three clonal variants which have reacquired EGF-mediated growth inhibition have 2- to 4-fold more EGF receptor than their respective parent variant. A biphasic response with stimulation at low and inhibition at high concentrations of EGF was especially evident in revertants of clone 29. Three separate antagonist monoclonal anti-EGF receptor antibodies block the growth inhibitory effects of EGF and uncover EGF-mediated growth stimulation. These studies indicate that in A431 cell variants a continuum of ligand-activated EGF receptors determines proliferative responses from low concentrations of active receptors under basal conditions to intermediate concentrations causing growth stimulation to high concentrations, causing inhibition of cell proliferation.
Growth stimulation of A431 cells by epidermal growth factor: identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Kawamoto, T, et al. Proc. Natl. Acad. Sci. U.S.A., 80: 1337-41 (1983)
1982
Epidermal growth factor (EGF) at 3 nM maximally inhibits the proliferation of A431 epidermoid carcinoma cells. We show that at lower concentrations, in the range of 3-100 pM, EGF has a mitogenic effect on A431 cells. In the presence of 100 nM anti-EGF-receptor monoclonal IgG (designated 528), which inhibits A431 cell proliferation and blocks greater than 95% of EGF binding, EGF becomes mitogenic for A431 cells at concentrations up to 3 nM. These results suggest that a minor population of high-affinity EGF receptors may be involved in stimulation of A431 cell proliferation. Saturation binding assays with 125I-labeled EGF indicate that approximately equal to 0.1-0.2% of receptors for EGF are high-affinity receptors that bind EGF with an estimated Kd of 7 X 10(-11) M. This affinity is nearly 2 orders of magnitude higher than that of the remaining EGF receptors. Although A431 cell proliferation is maximally inhibited by nonsaturating amounts of EGF (3 nM), maximal inhibition by 528 IgG (approximately equal to 70% of maximal inhibition by EGF) requires saturating concentrations of antibody (approximately equal to 15 nM). Unlike EGF, rapid down-regulation is not observed with 528 IgG. These results indicate different mechanisms of growth inhibition of A431 cells by EGF and 528 IgG.
