Abstract
Cyclic adenosine monophosphate (cAMP) is a second messenger of many G-protein-coupled receptors. The change in cellular cAMP level has widely been used to estimate the biological activity of various GPCR-specific agents. Förster resonance energy transfer (FRET) biosensors have been around for almost 10 years and became increasingly popular for cAMP detection. Ratiometric sensitized emission assay of a FRET biosensor can easily be implemented on fluorescence plate reader platforms. For such assays a considerable amount of cells expressing the desired biosensor is needed. A method to achieve sufficient and reproducible level of cAMP biosensor protein expression with the means of BacMam transduction system is the subject of this chapter.
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Mazina, O., Allikalt, A., Heinloo, A., Reinart-Okugbeni, R., Kopanchuk, S., & Rinken, A. (2015). cAMP assay for GPCR ligand characterization: Application of BacMam expression system. In G Protein-Coupled Receptor Screening Assays: Methods and Protocols (pp. 65–77). Springer New York. https://doi.org/10.1007/978-1-4939-2336-6_5
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