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Assay Method Information |
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| | Radioactive Kinase Assay |
| Description: | The assay was carried out in the reaction buffer containing Cdc7/Dbf4, His6-MCM2, and ATP/[gamma-33P]ATP in the presence of test compounds. After incubation, SPA (scintillation proximity assay) beads coated with anti-His6 antibody (Amersham) were added for an additional 1 h-incubation. The beads were collected by vacuum filtration, washed, and counted in a Microbeta reader (PerkinElmer Wallac). The percent MCM2 phosphorylation was calculated by taking into account the specific activity of the [gamma-33P]ATP and 60% counting efficiency. |
| Affinity data for this assay | |
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