| Chapter title |
Detection and Quantification of Intracellular Signaling Using FRET-Based Biosensors and High Content Imaging
|
|---|---|
| Chapter number | 10 |
| Book title |
G Protein-Coupled Receptors in Drug Discovery
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2914-6_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2913-9, 978-1-4939-2914-6
|
| Authors |
Michelle L. Halls, Daniel P. Poole, Andrew M. Ellisdon, Cameron J. Nowell, Meritxell Canals, Halls, Michelle L., Poole, Daniel P., Ellisdon, Andrew M., Nowell, Cameron J., Canals, Meritxell |
| Abstract |
Förster resonance energy transfer (FRET) biosensors represent invaluable tools to detect the spatiotemporal context of second messenger production and intracellular signaling that cannot be attained using traditional methods. Here, we describe a detailed protocol for the use of high content imaging in combination with FRET biosensors to assess second messenger production and intracellular signaling in a time-effective manner. We use four different FRET biosensors to measure cAMP levels, kinase (ERK and PKC), and GTPase activity. Importantly, we provide the protocols to express and measure these sensors in a variety of model cell lines and primary dorsal root ganglia neurons. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 14 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 3 | 21% |
| Researcher | 3 | 21% |
| Lecturer | 1 | 7% |
| Student > Ph. D. Student | 1 | 7% |
| Student > Doctoral Student | 1 | 7% |
| Other | 2 | 14% |
| Unknown | 3 | 21% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 3 | 21% |
| Pharmacology, Toxicology and Pharmaceutical Science | 2 | 14% |
| Agricultural and Biological Sciences | 2 | 14% |
| Biochemistry, Genetics and Molecular Biology | 1 | 7% |
| Neuroscience | 1 | 7% |
| Other | 1 | 7% |
| Unknown | 4 | 29% |