New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
Protein-based, fluorescent biosensors power basic research on cell signaling in health and disease, but their use in automated laboratories is limited. We have now created two live-cell assays, one for diacyl glycerol and another for cAMP, that are robust (Z′ > 0.7) and easily deployed on standar...
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| Published in | Journal of biomolecular screening Vol. 21; no. 3; pp. 298 - 305 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Los Angeles, CA
SAGE Publications
01.03.2016
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| Subjects | |
| Online Access | Get full text |
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| Abstract | Protein-based, fluorescent biosensors power basic research on cell signaling in health and disease, but their use in automated laboratories is limited. We have now created two live-cell assays, one for diacyl glycerol and another for cAMP, that are robust (Z′ > 0.7) and easily deployed on standard fluorescence plate readers. We describe the development of these assays, focusing on the parameters that were critical for optimization, in the hopes that the lessons learned can be generalized to the development of new biosensor-based assays. |
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| AbstractList | Protein-based, fluorescent biosensors power basic research on cell signaling in health and disease, but their use in automated laboratories is limited. We have now created two live-cell assays, one for diacyl glycerol and another for cAMP, that are robust (Z′ > 0.7) and easily deployed on standard fluorescence plate readers. We describe the development of these assays, focusing on the parameters that were critical for optimization, in the hopes that the lessons learned can be generalized to the development of new biosensor-based assays. |
| Author | Hughes, Thomas E. Shaner, Nathan C. Martinka, Scott Quinn, Anne Marie Tewson, Paul H. |
| Author_xml | – sequence: 1 givenname: Paul H. surname: Tewson fullname: Tewson, Paul H. – sequence: 2 givenname: Scott surname: Martinka fullname: Martinka, Scott – sequence: 3 givenname: Nathan C. surname: Shaner fullname: Shaner, Nathan C. – sequence: 4 givenname: Thomas E. surname: Hughes fullname: Hughes, Thomas E. – sequence: 5 givenname: Anne Marie surname: Quinn fullname: Quinn, Anne Marie |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/26657040$$D View this record in MEDLINE/PubMed |
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| Keywords | GFP diacyl glycerol baculovirus fluorescence adenylyl cyclase live-cell assay DAG mNeonGreen cAMP Z′ statistic |
| Language | English |
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| SubjectTerms | Automation, Laboratory Baculoviridae - physiology Biosensing Techniques Cyclic AMP - chemistry Cyclic AMP - metabolism Diglycerides - chemistry Diglycerides - metabolism HEK293 Cells Humans Reproducibility of Results Transduction, Genetic |
| Title | New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories |
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