| Assay Method Information | |
| | Autotaxin (ATX) Enzymatic Activity Inhibition Assay |
| Description: | The inhibitory activity of the compound against the Autotaxin enzyme was detected using the Autotaxin Inhibitor Screening Assay Kit (Cayman, 700580). The compound to be tested was first prepared into 10 mM stock solution in DMSO solvent, and then serially diluted into 8 concentrations of compound with DMSO. Subsequently, 8 concentrations of compound were diluted to 19× compound working solution (with DMSO content of 1.9%) with an Autotaxin Assay buffer (1×) provided in the kit. Autotaxin Assay Reagent (10×) was taken out and diluted 10-fold with Autotaxin Assay Buffer (1×). The Autotaxin Substrate was taken out, and added with 1.2 mL of Autotaxin Assay Buffer (1×) for dissolving. The mixture was mixed well and then let stand at room temperature. In a 96-well plate, 150 μL of Autotaxin Assay Buffer (1×), 10 μL of diluted 19× compound working solution, 10 μL of Autotaxin Assay Reagent (1×), 20 μL of dissolved Autotaxin Substrate were added into wells at each concentration, and then the mixture was mixed well, incubated at 37° C. on a constant-temperature shaking shaker, and incubated in the dark for 30 min. |
| Affinity data for this assay | |
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