| Assay Method Information | |
| | Ataxia-Telangiectasia Mutated (ATM) Biochemical Assay Using Reaction Biology's HTRF Kinase Assay Protocol |
| Description: | The assay detects kinase activity by antibody based HTRF assay. The GST-tagged p53 protein serves as substrate and Eu-labeled-antibody with specific phosphorylation site binds upon phosphorylation which is monitored by HTRF from the anti-GST-d2.Reagent: Reaction buffer; 50 mM Hepes (pH 7.5), 150 mM NaCl, 10 mM MnCl2, 1% Glycerol, 0.01% Brij35, 1 mM DTT, 1% DMSO.Substrate: Human p53 (aa2-393), RBC produced in E. coli, N-terminal GST-Tag, MW=70.42 kDa.[0429]Detection; MAb Anti-p53-pS15-Eu cryptate, CisBio cat #61P08KAZ; MAb Anti GST-d2Reaction Procedure: 1. Deliver 2× Enzyme in wells of reaction plate except No E control wells. Add buffer instead; 2. Deliver compounds in 100% DMSO into the kinase reaction mixture by Acoustic technology (Echo550; nanoliter range), incubate for 30 min at room temperature; 3. Deliver 2× substrate+ATP mixture into the reaction mixture to initiate the reaction; 4. Incubate for 2 hours at room temperature. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |