| Solution Information | help | |
| Enzyme: | Transitional endoplasmic reticulum ATPase | |
| inhibitor: | BDBM46552 | |
| 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 dose response curves for 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). This biochemical assay employs 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 p97 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 a nominal concentration of 50 uM. Protocol Summary: Prior to the start of the assay 18 ul of Assa | |
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