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Assay Method Information |
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| | Biological Assay |
| Description: | Kinase reaction buffer for p38alpha HTRF assays consists of 50 mM Tris-pH 7.5, 5 mM MgCl2, 0.1 mg/mL BSA, 100 uM Na3VO4 and 0.5 mM DTT. HTRF Detection Buffer:HTRF detection buffer contains 100 mM HEPES-pH 7.5, 100 mM NaCl, 0.1% BSA, 0.05% Tween-20, and 10 mM EDTA. Serial Dilution of Compounds: Compounds were dissolved in 100% DMSO and serially diluted (3 fold, 10 point) in a polypropylene 96-well microtiter plate (drug plate). The final starting concentration of compounds in the p38alpha enzymatic assays was 1 uM. Columns 6 and 12 (HI controls and LO controls respectively) in the drug plate were reserved as controls and contained only DMSO.Kinase Reaction: The p38alpha kinase reactions were carried out in a polypropylene 96-well black round bottom assay plate in total volume of 30 uL kinase reaction buffer. |
| Affinity data for this assay | |
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