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| | Fluorescence Polarization-based Assay |
| Description: | In this assay, a fluorescent phosphopeptide tracer and the nonfluorescent phosphopeptides generated during a PKA reaction compete for binding to an antiphosphoserine antibody. In a reaction mixture containing phosphopeptide product, the fluorescent tracer is displaced from the antibody and the emission signal becomes depolarized. Enzymatic activity was measured by detection of fluorescence polarity in a Victor 2 reader (Wallac, Finland). To determine IC50 values, the compounds were tested in the concentration range from 100 uM to 20 pM. Calculation of IC50 values was performed with ActivityBase. |
| Affinity data for this assay | |
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