| Assay Method Information | |
| | Homogeneous Time-Resolved Fluorescence (HTRF) Assay |
| Description: | The assay uses purified enzyme interacting with biotinylated peptide substrate.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. Relative fluorescence units were read on a Discovery plate reader (Packard). The IC50 values for inhibitors were determined by a four-parameter sigmoidal curve fit. |
| Affinity data for this assay | |
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