Development of a Fluorescent Quenching Based High Throughput Assay to Screen for Calcineurin Inhibitors
Fig 3
White plate fluorescence assay was validated using CaN and a known inhibitor. (a) Complete enzyme assay was performed in 272 replicates (full reaction) and compared with 80 replicates of no enzyme control. Assay was done manually in 384 well plates. The assay yielded a Z score of 0.63. (b) Endothall, a known inhibitor of CaN, was used to validate the assay format. The assay was done in duplicate and the data point represents mean and standard error. IC50 was calculated by prism software, using Eq 5, was 12.63μM.