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...

Descripción completa

Detalles Bibliográficos
Autores principales: Tewson, Paul H., Martinka, Scott, Shaner, Nathan C., Hughes, Thomas E., Quinn, Anne Marie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2015
Materias:
Original Research
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4766961/
https://www.ncbi.nlm.nih.gov/pubmed/26657040
http://dx.doi.org/10.1177/1087057115618608
_version_ 1782417761116356608
author Tewson, Paul H.
Martinka, Scott
Shaner, Nathan C.
Hughes, Thomas E.
Quinn, Anne Marie
author_facet Tewson, Paul H.
Martinka, Scott
Shaner, Nathan C.
Hughes, Thomas E.
Quinn, Anne Marie
author_sort Tewson, Paul H.
collection PubMed
description 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.
format Online
Article
Text
id pubmed-4766961
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher SAGE Publications
record_format MEDLINE/PubMed
spelling pubmed-47669612016-04-01 New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories Tewson, Paul H. Martinka, Scott Shaner, Nathan C. Hughes, Thomas E. Quinn, Anne Marie J Biomol Screen Original Research 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. SAGE Publications 2015-12-11 2016-03 /pmc/articles/PMC4766961/ /pubmed/26657040 http://dx.doi.org/10.1177/1087057115618608 Text en © 2015 Society for Laboratory Automation and Screening http://creativecommons.org/licenses/by-nc/3.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 3.0 License (http://www.creativecommons.org/licenses/by-nc/3.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Research
Tewson, Paul H.
Martinka, Scott
Shaner, Nathan C.
Hughes, Thomas E.
Quinn, Anne Marie
New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
title New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
title_full New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
title_fullStr New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
title_full_unstemmed New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
title_short New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories
title_sort new dag and camp sensors optimized for live-cell assays in automated laboratories
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4766961/
https://www.ncbi.nlm.nih.gov/pubmed/26657040
http://dx.doi.org/10.1177/1087057115618608
work_keys_str_mv AT tewsonpaulh newdagandcampsensorsoptimizedforlivecellassaysinautomatedlaboratories
AT martinkascott newdagandcampsensorsoptimizedforlivecellassaysinautomatedlaboratories
AT shanernathanc newdagandcampsensorsoptimizedforlivecellassaysinautomatedlaboratories
AT hughesthomase newdagandcampsensorsoptimizedforlivecellassaysinautomatedlaboratories
AT quinnannemarie newdagandcampsensorsoptimizedforlivecellassaysinautomatedlaboratories

Ejemplares similares