Recombinant Human EGFR Protein (672-1210, L858R), FLAG-tagged
Product Description
Cat
IMP-6458
Official Symbol
EGFR
Product Overview
Recombinant human EGFR (672-1210, L858R) protein that includes amino acids 672-1210 of human EGFR protein (accession number NP_005219.2) with a point mutation Leu858Arg, was expressed in baculovirus expression system with an N-terminal FLAG-Tag. The molecular weight of this protein is 63.1 kDa.
Description
EGFR (Epidermal Growth Factor Receptor), also known as ERBB, mENA, ERBB1 and HER1, is the receptor for members of the EGF family and is a transmembrane glycoprotein that has tyrosine kinase activity. Receptor 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, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homoand/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades, including at least 4 major downstream signaling cascades: the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules, leading to cell proliferation. EGFR is widely recognized for its importance in cancer. Amplification and mutations of this gene have been shown to be driving events in many cancer types. Its role in non-small cell lung cancer, glioblastoma and basal-like breast cancers has spurred many research and drug development efforts. In particular, EGFR mutation analysis of NSCLC (non-small cell lung cancer) is now routine in standard clinical practice, and tyrosine kinase inhibitors (TKIs) targeting EGFR-activating mutations are the most widely used targeted therapy, most notably gefitinib and erlotinib. Patients carrying a point mutation in exon 21 (L858R) or a deletion in exon 19, which account for approximately 90% of EGFR-activating mutations, have significant survival benefit when treated with EGFR-TKI.
Expression System
Insect cells
Species
Human
Tag
FLAG
Form
Recombinant EGFR (672-1210, L858R) protein is supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.
Molecular Mass
63.1 kDa
Protein length
672-1210
Purity
>52%
Applications
Enzyme kinetics, inhibitor screening, and selectivity profiling
Storage
Recombinant proteins in solution are temperature sensitive and must be stored at -80 centigrade to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
SDS-PAGE
SDS-PAGE

Recombinant EGFR (672-1210, L858R) protein 10% SDS-PAGE Coomassie staining MW: 63.1 kDa Purity: >52%

Bioactivity-ELISA 1
Activity 1

HTRF assay for EGFR (672-1210, L858R) protein activity 1 μM TK substrate was incubated with different concentrations of EGFR (672-1210, L858R) protein in a 10 μL reaction system containing 1×Enzymatic Buffer, 5 mM MgCl2, 1 mM DTT, 5 nM SEB and 100 uM ATP for 1 hour. Then 10 μL detection reagents containing TK antibody and SA-XL665 (each of which was 1:100 diluted with 1× Detection Buffer) were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature. HTRF KinEASE TK assay was used to detect the enzymatic activity.

Bioactivity-ELISA 2
Activity 2

HTRF assay for EGFR (672-1210, L858R) protein activity 1 μM TK substrate was incubated with different concentrations of EGFR (672-1210, L858R) protein in a 10 μL reaction system containing 1×Enzymatic Buffer, 5 mM MgCl2, 1 mM DTT, 5 nM SEB and 100 uM ATP for 1 hour. Then 10 μL detection reagents containing TK antibody and SA-XL665 (each of which was 1:100 diluted with 1× Detection Buffer) were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature. HTRF KinEASE TK assay was used to detect the enzymatic activity.

Data Sheet MSDS
For Research Use Only. Not intended for any clinical use. No products from Creative BioMart may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative BioMart.

Online Inquiry
Contact Info