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. Author manuscript; available in PMC: 2010 Nov 29.
Published in final edited form as: J Med Chem. 2010 Jan 14;53(1):37–51. doi: 10.1021/jm901070c

Figure 5.

Figure 5

Fluorescence Interference Analysis. A) Plate heatmaps associated with one 7-concentration compound series in 1,536-well format. Shown are the first and last fluorescent reads and the activity calculated from the 60-second initial rates. Subtracting out the higher-than-average but steady fluorescence of some compounds (red dots on heatmaps associated with first and last reads) leads to the significant reduction of interference (small number of blue dots in the activity heatmap). However, for compound 42 which is highly fluorescent (left plots in B and C), the drift in inherent fluorescence within the reaction time course (indicated within panel B) can lead to the erroneous calculation of concentration-response effect (C, left plot). A non-fluorescent inactive compound 43 (right plots in B and C) displays nearly overlapping reaction time courses (B, right plot) and as a consequence the corresponding concentration-response trend is relatively flat, resulting in the correct assignment of inactive phenotype (C, right plot).