| Assay Method Information | |
| | Kinase Inhibitory Assay |
| Description: | Table 32: The reagent used was as follows: Base Reaction buffer; 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.016 Brij35, 0.02 mg/ml BSA, 0.1 mM NaVO4, 2 mM DTT, 10 DMSO. Required cofactors were added individually to each kinase reaction.The reaction procedure was as follows:1) Substrates were prepared in freshly prepared Reaction Buffer.2) Any required cofactors were delivered to the substrate solution above.3) Kinase was delivered into the substrate solution and gently mixed.4) Compounds were delivered in 100% DMSO into the kinase reaction mixture by Acoustic technology (Echo550; nanoliter range), followed by incubation for 20 min at room temp.5) 33P-ATP was delivered into the reaction mixture to initiate the reaction.6) The mixture was incubated for 2 hours at room temperature.7) Kinase activity was detected by P81 filter-binding method. |
| Affinity data for this assay | |
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