Assay Method Information

Assay Name:  Homogeneous Time-Resolved Fluorescence (HTRF) Enzyme Assay
Description:  The assay uses purified GST-Tie-2 or GST-VEGFR2 interacting with biotinylated peptide substrates. HTRF is based on the proximity of europium cryptate (donor fluorophore)-labeled antiphosphotyrosine and streptavidin labeled with APC (allophycocyanin), which have been brought together by a binding reaction. When the two entities come into close proximity and upon excitation, energy transfer occurs and APC re-emits at 665 nm. The IC50 values for inhibitors were determined by a four-parameter sigmoidal curve fit.
Affinity data for this assay
 

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