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| | HTRF Kinase Inhibition Assay |
| Description: | In vitro kinase assays were done to establish IC50 values against recombinant enzymes using homogeneous time-resolved fluorescence (HTRF) assay. For all assays, streptavidin-allophycocyanin and Eu-PT66 were added immediately before the HTRF reaction. Molecules were tested in a 10-point serial dilution using an ATP concentration of two thirds Km values that were determined for the enzymes and calculated using the Eadie-Hofstee and Lineweaver-Burke methods. The fluorescence ratios were read on a Rubystar instrument (BMG labtech Inc.). |
| Affinity data for this assay | |
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