| Solution Information | help | |
| Enzyme: | Hexokinase-1 | |
| inhibitor: | BDBM80774 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | A. Brief Description of the Assay: This assay determined the potency of activators or inhibitors of hexokinase I (HK1) enzyme. It is measured via fluorescence in 1536 well plate format. B. Materials: Item, Source, Cat# HK1 Enzyme Stock Solutions, In-house, N/A Glucose, Sigma-Aldrich, G6152 NADP, Amresco, 0760 Resazurin, Sigma-Aldrich, R7017 ATP, Sigma-Aldrich, A7699 Diaphorase, Sigma-Aldrich, D5540 G6PDH, Sigma-Aldrich, G8404 Tris-HCl pH 8.0, Sigma-Aldrich, T6066 KCl, Sigma-Aldrich, P9541 Tween-20, Sigma-Aldrich, P1379 MgCl2, Sigma-Aldrich, M8266 Bovine serum albumin, Sigma-Aldrich, A7888 Beta-mercaptoethanol, Calbiochem, 444203 Mol. Grade H2O, Mediatech, 46-000-CM 1536-well, black, non-binding plates, Aurora, 00029844 C. Procedures: 1. Prepare Reagents as described in section D. Recipe. 2. Using LabCyte Echo, transfer varying volumes of test compound in DMSO from Echo qualified plate into assay plate Col. 5 - 48 to achieve the desired dose response concentrations and range. Backfil | |
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