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Assay Method Information

Assay Name:  Homogeneous Time-Resolved Fluorescence (HTRF) Enzyme Assay
Description:  The assay uses purified enzyme interacting with biotinylated peptide substrate. HTRF is based on the proximity of europium cryptate (donor fluorophore)-labeled a ntiphosphotyrosine 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. Relative fluorescence units were read on a Ruby Star fluorescent reader (BMG Technologies, Inc.) using a four-parameter fit using activity base to calculate the corresponding IC50 for the test and standard compounds in each well.
Affinity data for this assay
 

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Last update November 1, 2007
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