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Use of BRET2 luminescence cell-based assays with LEADseeker Multimodality Imaging System

assay uses the luminescence emission signal from the donor and the luminescence energy transfer signal from the acceptor to generate the two sets of data used for the final analysis. The filters developed for the system are configured for donor emission at 470 nm and 535 nm for acceptor emission.

Table 1 shows the raw signal data generated on the LEADseeker Multimodality Imaging System for the donor and acceptor channels. Signal to noise was calculated using the following equation:

S:N values for all conditions in both assay volumes was < 10:1 (comparable with pack leaflet data), indicating that the signals generated were sufficient for final data analysis.

In the final data analysis, we ratioed the data obtained from the two channels of the raw signal analysis experiment and compared the ratios for the positive and negative control cell extracts (Fig 1).



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