Quantification of the effects of mutations on receptor tyrosine kinase (RTK) activation in mammalian cells.

Single amino acid mutations in receptor tyrosine kinases (RTKs) are known to cause receptor over-activation and disease. Here we present a detailed protocol for the quantification of the effect of mutations on RTK activation in mammalian cells. The activation measurements are based on Western blotting, and involve direct comparison of receptor phosphorylation under conditions that ensure identical expression of wild-type and mutant receptors.