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Receptor Binding Assays using Fluorescence Polarization Detection: the [FP]2TM Characterized Tracers

ontains performance data obtained in a similar manner to Vasopressin for all the peptides listed in Table 1. In most cases, the assay window, defined as the difference between the Bound and Displaced signals shown in Figure 4 (Δ mP), is either approaching 100 mP or exceeding it. Furthermore, each of the assays demonstrates a Z factor > 0.5, indicating "an excellent assay" which can easily transition to a high throughput screen.

Summary

All fluorescently-labeled peptides listed in Tables 3 can be used to perform high throughput primary screening of compounds using fluorescence polarization detection. Appropriate pharmacology and satisfactory assay performance enables an easy transition to screening conditions. This simple and robust assay platform allows for ultra high throughput screening for GPCR binding applications.

References

1. Divers, M. (1999). What is the future of high throughput screening? J. Biomol. Screen. 4:177-178.

2. Eglen, R. M. (1999). High throughput screening: myths and future realities. J. Biomol. Screen. 4:179-181.

3. High-Throughput Screening 2000: New Trends and Directions (2000). HighTech Business Decisions , Moraga, CA.

4. Allen, M., Reeves, J., Mellor, G. (2000). High throughput fluorescence polarization: a homogeneous alternative to radioligand binding for cell surface receptors. J. Biomol. Screen. 5:63-70.

5. Banks, P., Gosselin, M., Prystay, L. (2000). Fluorescence polarization assays for high throughput screening of G protein-coupled receptors. J. Biomol. Screen. 5:159-168.

6. Lee, P. Bevis, D. (2000). Development of a homogeneous high throughput fluorescence polarization assay for G protein-coupled receptor binding. J. Biomol. Screen. 5:415-419.

7. Turconi, S., Shea, K., Ashman, S. et al. (2001). Real experiences of
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