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Kinase Assay Protocol
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发布时间：2024-05-10
Base Reaction buffer; 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO*Required cofactors are added individually to each kinase reactionReaction Procedure: Prepare indicated substrate in freshly prepared Base Reaction BufferDeliver any required cofactors to the substrate solution aboveDeliver indicated kinase into the substrate solution and gently mixDeliver compounds in DMSO into the kinase reaction mixture by Acoustic technology (Echo550; nanoliter range), inclubate for 20 minutes at room temperatureDeliver 33P-ATP (specific activity 10 Ci/l) into the reaction mixture to initiate the reaction.Incubate kinase reaction for 2 hours at room temperatureReactions are spotted onto P81 ion exchange paper Detect kinase activity by filter-binding method.>>Return to Kinase Page
本文链接： http://reaction.immuno-online.com/view-1497844892.html
发布于 ： 2024-05-10
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