Titlebook: cAMP Signaling; Methods and Protocol Manuela Zaccolo Book 2015 Springer Science+Business Media New York 2015 Adenosine 3′,5′-monophosphate.

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1064-3745 chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting an978-1-4939-4950-2978-1-4939-2537-7Series ISSN 1064-3745 Series E-ISSN 1940-6029

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https://doi.org/10.1007/978-3-031-22210-8lls stimulated with glucose and gluco-incretin hormones. We also demonstrate how the technique can be combined with measurements of the cytosolic Ca. concentration or with recordings of the subcellular localization of the cAMP effector protein Epac2. The translocation reporter approach provides a va

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Applications and Usability of Interactive TViovascular, neuronal, and inflammatory diseases, but it is unclear whether these interactions are suitable drug targets. Here we describe an enzyme-linked immunosorbent assay (ELISA) for the screening of small molecule libraries for inhibitors of AKAP–PKA interactions. In addition, we describe a hom

FRET

Marc Aguilar,Pau Pamplona,Sergi FernándezPKG and their associated proteins. Unfortunately, both PKA and PKG are enriched in the pull downs with both immobilized compounds. Although this proved sufficient to identify novel AKAPs, the lower abundance of PKG has seriously hampered the enrichment and identification of novel GKAPs. Here we pres

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Adenoviral Transduction of FRET-Based Biosensors for cAMP in Primary Adult Mouse Cardiomyocytes,ts a powerful combination for the study of cAMP signaling in live primary cardiomyocytes. In this chapter, we describe the steps required during the isolation, viral transduction, and culture of cardiomyocytes from an adult mouse to obtain reliable expression of genetically encoded FRET biosensors for the study of cAMP signaling in living cells.

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Photoactivatable Adenylyl Cyclases (PACs) as a Tool to Study cAMP Signaling In Vivo: An Overview,ols provide cyclase activity capable of increasing cellular cAMP levels up to a hundredfold with either . or . kinetic characteristics. Here, we consider the functional features of different cyclases and provide operating guidelines to optimize the use of PACs in vivo.

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