| Assay Method Information | |
| | CMP Assay |
| Description: | The assay detects the product (free CoA) which is produced from palmitoyl-CoA as a method to detect the enzymatic activity. Test compounds that inhibit TEAD activity are therefore able to block the CMP fluorescent signal.TEAD2/4 protein (10 μL of a 50 ng/μL solution) and a solution the test compound (0.5 μL, variable concentration) were pre-incubated for 30 min at r.t. A mixture prepared from MES pH 6.4 buffer (6.5 μL of a 50 mM solution), EDTA (1 μL of a 20 mM solution), palmitoyl-CoA (1 μL of a 20 μM solution), and 7-diethylamino-3-(4′-maleimidylphenyl)-4-methylcoumarin (1 μL of a 10 μM solution) was added and the resulting mixture was mixed well using a pipette. The solution was then incubated at r.t. for 30-120 min. in the dark, and fluorescence was detected at 350 nm every 30 min. until the fluorescence signal is saturated. An IC50 for the ability of the compound to inhibit fluorescence is calculated from the results. |
| Affinity data for this assay | |
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