| Assay Method Information | |
| | Measurement of Mat2A inhibition is performed in 384 well format absorbance-based assay |
| Description: | Recombinant human Mat2a (12.5 nM) and serial diluted compounds in DMSO (range of concentrations from 10 μM to 508 μM) or controls (DMSO) are incubated for 15 minutes at room temperature (RT) in assay buffer containing 50 mM HEPES pH 7.5, 50 mM KCl, 50 mM MgCl2, 0.01% Tween 20 and 10 mM DTT. The reaction is initiated by the addition of the combined substrates ATP and Methionine, each at a final concentration of 100 μM. Final assay condition are 12.5 nM Mat2A, 100 μM ATP and Methionine Substrates and 2% DMSO. After 120 minutes of incubation at RT, the reaction is stopped by the addition of Biomol Green. The absorbance signal is measured at λ=635 nm with a multiplate reader (BMG Pherastar reader or equivalent) after 30 min of equilibration at RT. |
| Affinity data for this assay | |
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