Recombinant human epidermal growth factor receptor (EGFR), protein kinase domain, 10 µg

Background: The epidermal growth factor receptor (EGFR) is part of the erbB family of receptor tyrosine kinases, including also HER2/erbB2, HER3/erbB3 and HER4/erbB4. Activation of ErbB receptors is initiated by ligand binding to their extracellular domain, leading to the formation of both, homo- and heterodimeric ErbB receptor complexes. Upon dimerization the receptor kinase undergo autophosphorylation at specific tyrosine residues within the intracellular domain, acting as binding sites for proteins e.g. Shc, Grb2, Src, PI3K,  SHP1 and SHP2.  This couples the ErbB receptor to downstream signaling pathways e.g. MAPK and the Akt pathway, resulting in the activation of target genes within the nucleus, which determine biological responses, such as proliferation or differentiation. Deregulated EGFR signaling, caused by overexpression of the receptor or an enhanced ligand production, is frequent in many tumor types. Thus, there is a great interest in generation of specific inhibitors. Gefitinib (Iressa) is an orally available ATP competitive EGFR-inhibitor which had demonstrated significant efficacy in non-small cell lung cancer. 

Protein: human EGF-R kinase domain (amino acids H672-A1210, GenBank entry NM_ 005228),  contains N-terminal GST-His6 fusion tags with a 3C cleavage sites
Theoretical MW (EGF-R): 89.2 kDa (fusion protein)
Expression system: Baculovirus infected Sf9 cells
Purification: One-step affinity purification using glutathione agarose
Storage buffer: 50 mM Hepes pH 7.5, 100 mM NaCl, 5 mM DTT, 15 mM reduced glutathione, 20 & glycerol
Protein concentration: 0.468 mg/ml (Bradford method using BSA as standard protein)
Method for determination of Km value & specific activity: Filter binding assay MSFC membrane
Specific activity : 388,000 pmol/mg min

Entrez Gene ID: 1956
UniProtKB:  P00533

BIAFFIN – SERVICES:

Use our EGFR - SPR Binding Assay for comprehensive kinetic characterization of your small molecule kinase inhibitors. 

Figure: Real-time kinetic analysis of kinase inhibitor Erlotinib binding to EGFR using surface plasmon resonance.

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Product specific literature references:

Ono M, Kuwano M.(2006) " Molecular mechanisms of epidermal growth factor receptor (EGFR) activation and response to gefitinib and other EGFR-targeting drugs." Clin Cancer Res, 12(24):7242-51.

Takayuki Fukui, MD, Tetsuya Mitsudomi, MD.(2008) " Mutations in the epidermal growth factor receptor gene and effects of EGFR-tyrosine kinase inhibitors on lung cancers". Gen Thorac Cardiovasc Surg. 56:97-103.

Uberall I, Kolïar Z, Trojanec R, Berkovcova J, Hajdïuch M.(2008) "The status and role of ErbB receptors in human cancer". Exp Mol Pathol. 84(2):79-89