| Assay Method Information | |
| | FAK Kinase Assay |
| Description: | The inhibitory activity of a compound against FAK was determined in a buffer containing 5 mM HEPES, pH 7.5, 0.01% Triton, 0.5 mM EGTA, and 0.01% BRIJ™-35 in a 384-well assay plate. The assay was carried out in the presence of the FAK at 1 nM and ATP at 6.8 μM using fluorescein-polyGT (0.2 μM) as an FAK substrate. Each assay mixture was incubated at room temperature for 30 min. A LANTHASCREEN® Tb-PY20 antibody was added to each well to stop the enzymatic reaction. The plate was incubated at room temperature for 60 min before reading on an ENVISION using TR-FRET module (excitation at 340 nm and emission at 520 nm and 495 nm). |
| Affinity data for this assay | |
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