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An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells
Here, we describe an optimized protocol to identify specific nuclear receptor ligands. First, to rule out any compound interference with luciferase activity per se, we describe an in vitro assay assessing potential inhibition or activation of luciferase enzymatic activity. Second, to comply with EMA...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8273406/ https://www.ncbi.nlm.nih.gov/pubmed/34286290 http://dx.doi.org/10.1016/j.xpro.2021.100658 |
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author | Berthier, Alexandre Staels, Bart Lefebvre, Philippe |
author_facet | Berthier, Alexandre Staels, Bart Lefebvre, Philippe |
author_sort | Berthier, Alexandre |
collection | PubMed |
description | Here, we describe an optimized protocol to identify specific nuclear receptor ligands. First, to rule out any compound interference with luciferase activity per se, we describe an in vitro assay assessing potential inhibition or activation of luciferase enzymatic activity. Second, to comply with EMA and FDA guidelines to mitigate drug-drug interactions, we detail assays assessing constitutive androstane receptor (CAR) and pregnane X receptor (PXR) activation ability. Finally, to minimize off-target detection effects, we describe the use of mammalian one- (or two-) hybrid systems. For complete details on the use and execution of this protocol, please refer to Hering et al. (2018). |
format | Online Article Text |
id | pubmed-8273406 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-82734062021-07-19 An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells Berthier, Alexandre Staels, Bart Lefebvre, Philippe STAR Protoc Protocol Here, we describe an optimized protocol to identify specific nuclear receptor ligands. First, to rule out any compound interference with luciferase activity per se, we describe an in vitro assay assessing potential inhibition or activation of luciferase enzymatic activity. Second, to comply with EMA and FDA guidelines to mitigate drug-drug interactions, we detail assays assessing constitutive androstane receptor (CAR) and pregnane X receptor (PXR) activation ability. Finally, to minimize off-target detection effects, we describe the use of mammalian one- (or two-) hybrid systems. For complete details on the use and execution of this protocol, please refer to Hering et al. (2018). Elsevier 2021-07-07 /pmc/articles/PMC8273406/ /pubmed/34286290 http://dx.doi.org/10.1016/j.xpro.2021.100658 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Berthier, Alexandre Staels, Bart Lefebvre, Philippe An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
title | An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
title_full | An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
title_fullStr | An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
title_full_unstemmed | An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
title_short | An optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
title_sort | optimized protocol with a stepwise approach to identify specific nuclear receptor ligands from cultured mammalian cells |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8273406/ https://www.ncbi.nlm.nih.gov/pubmed/34286290 http://dx.doi.org/10.1016/j.xpro.2021.100658 |
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