| Assay Method Information | |
| | In Vitro MTH1 Enzymatic Assay |
| Description: | Enzymatic reaction was initiated by adding either 8-oxo-dGTP (13.2 μm; TriLink BioTechnologies) or 2-OH-dATP (8.3 μm; Jena Bioscience) to reaction mixture containing test compounds, assay buffer [100 mm Tris-acetate, 40 mm NaCl, 10 mm Mg(OAc)2, 0.005% Tween-20, and 2 mm dithiothreitol (DTT), pH 7.5], His-taggedhuman recombinant MTH1 protein (2 nm; ProSpec), and mixed reagent from the PPiLight Inorganic Pyrophosphate Assay Kit (Lonza Rockland). The amount of pyrophosphate (PPi) generated as a result of MTH1 enzymatic activity was then monitored by recording luminescence signal for 30 min using a luminometer(Molecular Devices). |
| Affinity data for this assay | |
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