Solution Informationhelp
Enzyme: Wee1-like protein kinase
inhibitor: BDBM36838
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 cell-based high throughput screening assay for inhibitors of Wee1 degradation" (PubChem AID 1321), and that confirmed activity in a set of experiments entitled, "Confirmation cell-based high throughput screening assay for inhibitors of Wee1 degradation" (PubChem AID 1410). The assay uses HeLa cells transfected with a plasmid that encodes a Wee1K328M-luciferase fusion protein to monitor Wee1 levels. Wee1K328M is a kinase negative mutant used to avoid the cell growth inhibitory effects of wildtype wee1. The Wee1K328M-luciferase complex is rapidly turned over in these cells. As designed, compounds that inhibit Wee1 degradation will increase Wee1K328M-luciferase stability, leading to increased well luminescence. Compounds were tested in triplicate using a 10-point, 1:3 dilution series, starting at a nominal concentration of
 
 
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