Assay Method Information

Assay Name:  DNA-PK Biochemical Assay Using Reaction Biology's HotSpot Kinase Assay Protocol
Description:  Reagent: Base Reaction buffer; 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.01% Brij35, 0.02 mg/ml BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO Required cofactors are added individually to each kinase reaction.Reaction Procedure: 1. Prepare substrate in freshly prepared Reaction Buffer; 2. Deliver any required cofactors to the substrate solution above; 3. Deliver kinase into the substrate solution and gently mix; 4. Deliver compounds in 100% DMSO into the kinase reaction mixture by Acoustic technology (Echo550; nanoliter range), incubate for 20 min at room temperature; 5. Deliver 33P-ATP into the reaction mixture to initiate the reaction; 6. Incubate for 2 hours at room temperature; 7. Detect kinase activity by P81 filter-binding method.
Affinity data for this assay
 

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