Western Blotting Analysis as a Tool to Study Receptor Tyrosine Kinases
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Receptor tyrosine kinases (RTKs) are involved in critical aspects of cell physiology ranging from cell survival, proliferation, growth, migration, and differentiation. A tight control of the extent and duration of signals elicited by activated RTKs is crucial for preventing over-stimulation, which can ultimately lead to unrestrained proliferative ability and neoplastic growth. Ligand-induced downregulation of RTKs has emerged as a key negative regulatory mechanism that can accomplish signaling attenuation, by removing activated receptors from the cell surface and committing them to degradation. The ability of RTKs to escape from ligand-induced downregulation has been reported as a recurrent mechanism of oncogenic deregulation in cancer.
Western blotting procedures have been extensively proven as straightforward assays to evaluate protein expression levels and have been widely applied to study RTKs downregulation.
Key wordsProtein Western blotting Receptor tyrosine kinases
Preparation of this chapter was supported by the Health Research Board of Ireland (Grant RP/2006/77); Trinity College Dublin’s Start-Up Funds for New Academics 2008/2009 and Science Foundation Ireland’s funding of Molecular Therapeutics for Cancer, Ireland (08/SRC/B1410).
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