Acitve EGF Receptor EIA Kit





Product Name: Acitve EGF Receptor EIA Kit

# of Samples: 1X96 Assays

Intended Use: The Active EGF receptor EIA Kit is an in vitro enzyme immunoassay (EIA) kit for quantitative determination of human Phosphorylated EGF receptor in serum, cell lysate and other biological fluids.
This kit is for research use only. It is not for use in diagnostic or therapeutic procedures.

Introduction: The epidermal growth factor (EGF) receptor is a glycoprotein that is consist of 130 kDa protein (1186 amino acid residues) and 40 kDa sugar chain. EGF receptor molecule is made up of four domains; glycosylated extracellular domain that is the binding site of EGF and TGF?, a transmembrane domain, tyrosin kinase domain and auto-phosphorylation domain. Tyrosin kinase domain is an intracellular domain which is homologous with the oncogene v-erbB product. (1)
The binding of EGF to the receptor results in DNA reproduction and cell proliferation. During this step, ruffuring of cell membrane, phosphorylation of EGF receptor, internalization, pH change, enzyme activation, re-organization of actin filament, and oncogene protein induction are occured. Although the mechanism of the EGF signal transduction is not clear, it is assumed that EGF binding stimulation should initiate a series of signal transduction through tyrosine auto-phosphorylation and phosphorylation of interacting proteins.(2)
The half-life of EGF receptor on the cell surface is approximately 20 hours. When the receptor accepts the ligand, the degradation of the receptor is accelerated
(degraded approximately in 5 hours).(3)
The human squamous epithelium carcinoma cell line, A431, possess many EGF receptor, 5-10 times of standard cell line\'s. (2-3 x 106 / cell) This kit enables to measure the quantity of phosphorylated EGF receptor by ELISA. It is useful for analysis of receptor phosphorylation and for in vitro screening of specific inhibitors to EGF receptor kinase.

Principle of the test: The active EGF receptor EIA Kit is a solid phase EIA based on a sandwich method that utilizes mouse monoclonal anti human EGF receptor antibody and anti phosphotyrosine antibody to detect phosphrylated and activated EGF receptor by two-step procedure. The mouse monoclonal anti-human EGF receptor (B4G7) is immobilized onto the microtiter plate and blocked against non-specific binding. Samples and standards are added into each wells and incubated. The second step is to wash the plate and to add the second anti-phosphotyrosine (PY20) antibody labelled with peroxidase (POD). During this incubation, phosphorylated EGF receptor is bound to anti-EGF receptor (solid phase) on one side and tagged on the other by POD-anti-phosphotyrosin. The reaction between POD and substrate (H2O2, and 3,3\',5,5\'-tetramethylbenzidine) results in colour development with intensities proportional to the amount of phosphorylated (named active) EGF receptor present in samples and standards. The amount of active EGF receptor can be quantitated by measuring the absorbance using an EIA plate reader. Accurate sample concentrations of active EGF receptor can be determined by comparing their specific absorbances with those obtained for the standards plotted on a standard curve.

Storage and Stability: This kit is shipped at 2-8C and should be stored at 2-8 C if not used. Under this condition, the kit is stable until the expiry date printed in the box label.

Sensitivity and Specificity: This kit specifically measures phosphorylated EGF receptor. This kit can be used to measure only human phophorylated EGF receptor. Inactive EGF receptor that is dephosphorylated is not detected in this kit.

Product Name: Acitve EGF Receptor EIA Kit

Receptor for EGF, but also for other members of the EGF family, as TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor. Is involved in the control of cell growth and differentiation. Phosphorylates MUC1 in breast cancer cells and increases the interaction of MUC1 with C-SRC and CTNNB1/beta-catenin.

Function: Isoform 2/truncated isoform may act as an antagonist.

Catalytic Activity: ATP + a [protein]-L-tyrosine = ADP + a [protein]-L-tyrosine phosphate.

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. Binds MUC1.

Subcellular Location: Cell membrane; Single-pass type I membrane protein.

Subcellular Location: Isoform 2: Secreted.

Tissue Specificity: Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.

Ptm: Phosphorylation of Ser-695 is partial and occurs only if Thr-693 is phosphorylated.

Ptm: 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].

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.

Similarity: Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.

Similarity: Contains 1 protein kinase domain.

Additional Information

Name Acitve EGF Receptor EIA Kit
Related Product Names Acitve EGF Receptor EIA ELISA Kit; EC; Receptor tyrosine-protein kinase ErbB-1 ERBB1Acitve EGF Receptor EIA KitEGFR
Swissprot ID P00533
NCBI Acc Number NP_005219.2
Molecular Weight 134277
Datasheets/Manuals Printable datasheet for GWB-762CBD
Intended Use Research Use Only