| Assay Method Information | |
| | Biochemical Assay |
| Description: | The ADP-GLO Kinase Assay (Promega Corp., Madison, WI) measures ADP formed from a kinase reaction. According to the manufacture, the ADP formed in a kinase assay is first converted into ATP, which is then used to generate light in a luciferase reaction. The luminescence generated correlates with kinase activity. An exemplary experimental setting is described below, though minor adjustments can be made in individual assays. 1. Buffer Preparation40 mM Tris pH7.5; 20 mM MgC12, 0.1 mg/ml BSA, 50 µM DTT Buffer stock1 M Tris, PH7.5, 121.14 g/molAdd 6.057 g to 50 mL H2O, adjust PH to 7.51 M MgCl2, 95.21 g/molAdd 4.7605 g to 50 mL H2OAdd 20 mL 1 M Tris and 10 mL 1 M MgC12to 470 mL ddH2O to get buffer stock, and stock at RT.2. Preparing Fresh 1* Assay BufferReagent [Stock] [Final] Fold Add (mL)DTT (mM) 10 0.05 200 0.015BSA (mg/mL) 100 0.1 1000 0.003buffer stock 3 |
| Affinity data for this assay | |
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