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Assay Method Information |
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| | iFLik Assay |
| Description: | Labeled Akt1 was diluted to 200 nM in the measurement buffer consisting of 50 mMHEPES pH 7.4, 200 mM NaCl, and 0.01% Triton-X 100. Dilution series of candidate compounds were prepared in DMSO at 21× the final desired concentration. Compounds (0.5 μL) were then mixed with labeled Akt1 (10 μL) in triplicate and with measurement buffer alone (background fluorescence control) in quadruplicate at 12different concentrations. The final compound concentrations ranged from 0.8 nM to 500 μM and each well contained 5% v/v DMSO after mixing. Plates were then covered with an adhesive plastic foil and incubated for 2 h at RT prior to measurement of emission intensities at fluorophore-specific wavelengths |
| Affinity data for this assay | |
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