Patent ID: 7070938

Claim:
A method to measure the effect of a putative VEGF-R2 kinase activity modulating compound comprising the steps, in order: A) providing a test compound, a rat VEGF-R2 kinase fusion protein (SEQ ID NO:4) comprising N-terminal hexahistidine linked to amino acids 786–1343 of the rat VEGF-R2 protein in a solution suitable to provide rat VEGF-R2 catalytic activity and containing 33 P-γ-ATP as the source of phosphate; B) contacting the compound and the kinase fusion protein for sufficient time to provide a 33 P phosphorylated substrate; C) isolating the phosphorylated kinase fusion protein by affinity capture using a multiwell assay plate; D) removing remaining 33 P-γ-ATP by first aspirating the aqueous solution and then washing the plate with a phosphate buffered saline solution containing a divalent cation chelator in a concentration of about 1 mM to 100 mM; and E) detecting a change in kinase activity by monitoring the rate or absolute amount of 33 P transfer to the kinase fusion protein by autophosphorylation in the presence of the compound.