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In Vitro Enzyme Kinetics Analysis of EGFR

  • Zhihong WangEmail author
  • Christine Candelora
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1487)

Abstract

The epidermal growth factor receptor (EGFR) is an essential receptor tyrosine kinase (RTK) that regulates cell proliferation and differentiation, and its abnormal activation contributes to a variety of human cancers. EGFR is activated by the binding of growth factors, such as epidermal growth factor (EGF), resulting in the formation of an asymmetric dimer in which one EGFR molecule allosterically activates the other molecule. This chapter provides a detailed protocol to purify and characterize full-length EGFR bound with EGF. This protocol facilitates the expression and preparation of full-length EGFR in transiently transfected mammalian cell culture. A method is also presented for quantitative evaluation of EGFR enzyme activity.

Key words

Epidermal growth factor receptor Mammalian protein expression Transient transfection Kinase assay Radiometric assay 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Chemistry and BiochemistryUniversity of the SciencesPhiladelphiaUSA

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