| Solution Information | help | |
| Enzyme: | Transitional endoplasmic reticulum ATPase | |
| inhibitor: | BDBM46565 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine whether compounds identified as active in a previous set of experiments entitled, "Primary biochemical high-throughput screening assay to measure p97 ATPase inhibition" (PubChem AID 1481), and that confirmed activity in a subsequent set of experiments entitled, "Confirmation biochemical high-throughput screening assay for inhibitors of the p97 ATPase" (PubChem AID 1517) act as covalent or non-covalent p97 inhibitors. This biochemical assay employs a mutant Cys522Ala p97 protein as the target. The assay uses the Kinase-Glo reagent, which contains a luciferase that emits luminescence in direct proportion to ATP levels. As designed, compounds that inhibit the ATPase activity of p97Cys522Ala will reduce ATP hydrolysis, thereby increasing the relative levels of ATP available for consumption by luciferase, resulting in increased well luminescence. Compounds were tested in triplicate using a 10-point, 1:3 dilution series, starting at | |
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