Evaluating Opioid-Mediated Adenylyl Cyclase Inhibition in Live Cells Using a BRET-Based Assay
| Autor: | Preeti Manandhar, Marina Santiago, Shivani Sachdev |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Forskolin medicine.drug_class HEK 293 cells Drug action Transfection Cell biology Adenylyl cyclase 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine chemistry Opioid Opioid receptor medicine Receptor 030217 neurology & neurosurgery 030304 developmental biology medicine.drug |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071608838 |
| DOI: | 10.1007/978-1-0716-0884-5_11 |
| Popis: | Quantitative measurement of receptor signaling by different ligands is important for understanding the mechanism of drug action and screening of drugs. Here, we describe a simple and cost-effective method of measuring adenylyl cyclase inhibition, one of the hallmarks of opioid receptor activation. The assay is based on bioluminescence resonance energy transfer (BRET) that involves transfection of a biosensor in human embryonic kidney (HEK)-293 cells stably transfected with μ-opioid receptor (μ receptor), enabling real-time measurement of cAMP levels. |
| Databáze: | OpenAIRE |
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