| Assay Method Information | |
| | Evaluation of Inhibitory Activity of Enzyme Units |
| Description: | (3) Mlk3 (Human): MLK3 kinase protein and compounds were added to a buffer system containing 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.33 mg/mL myelin basic protein, 5 mM DTT, 10 mM Magnesium acetate and [γ-33P]-ATP, and then a Mg/ATP mixture was added to the system to initiate a reaction. After culturing at room temperature for 120 minutes, 0.5% phosphoric acid was added to terminate the reaction. 10 μL of the reaction mixture was spotted on a P30 filtermat and washed four times in 0.425% phosphoric acid for a total of 4 minutes. After washing once with methanol, the filtermat was dried and scintillated. |
| Affinity data for this assay | |
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