IMP-6456

EGFR

Recombinant human EGFR (672-1210) protein that includes amino acids 672-1210 of human EGFR protein (accession number NP_005219.2) was expressed in baculovirus expression system with an N-terminal FLAG tag. The molecular weight of this protein is 63.1 kDa.

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.

Insect cells

Human

FLAG

Recombinant EGFR (672-1210) 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.

63.1 kDa

672-1210

>55%

Enzyme kinetics, inhibitor screening, and selectivity profiling

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.

Recombinant EGFR (672-1210) protein gel 10% SDS-PAGE gel with Coomassie Blue staining MW: 63.1 kDa Purity: >55%

HTRF assay for EGFR (672-1210) protein activity 1 μM TK substrate was incubated with different concentrations of EGFR (672-1210) 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 hr. Then 10 ul 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 reactions were performed at room temp. HTRF assay was used for detection.