Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET

This protocol is used to profile the engagement of kinase inhibitors across nearly 200 kinases in a live-cell context. This protocol utilizes one single kinase tracer (NanoBRET(TM) Tracer K10) that operates quantitatively at four different concentrations. Minimizing the number of tracers offers a si...

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Autores principales: Robers, Matthew B., Wilkinson, Jennifer M., Vasta, James D., Berger, Lena M., Berger, Benedict-Tilman, Knapp, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449129/
https://www.ncbi.nlm.nih.gov/pubmed/34568844
http://dx.doi.org/10.1016/j.xpro.2021.100822
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author Robers, Matthew B.
Wilkinson, Jennifer M.
Vasta, James D.
Berger, Lena M.
Berger, Benedict-Tilman
Knapp, Stefan
author_facet Robers, Matthew B.
Wilkinson, Jennifer M.
Vasta, James D.
Berger, Lena M.
Berger, Benedict-Tilman
Knapp, Stefan
author_sort Robers, Matthew B.
collection PubMed
description This protocol is used to profile the engagement of kinase inhibitors across nearly 200 kinases in a live-cell context. This protocol utilizes one single kinase tracer (NanoBRET(TM) Tracer K10) that operates quantitatively at four different concentrations. Minimizing the number of tracers offers a significant workflow improvement over the previous protocol that utilized a combination of 6 tracers. Each NanoBRET(TM) kinase assay is built using commercially available plasmids and has been optimized for NanoLuc tagging orientation, diluent DNA, and tracer concentration. For complete details on the use and execution of this protocol, please refer to Vasta et al. (2018).
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spelling pubmed-84491292021-09-24 Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET Robers, Matthew B. Wilkinson, Jennifer M. Vasta, James D. Berger, Lena M. Berger, Benedict-Tilman Knapp, Stefan STAR Protoc Protocol This protocol is used to profile the engagement of kinase inhibitors across nearly 200 kinases in a live-cell context. This protocol utilizes one single kinase tracer (NanoBRET(TM) Tracer K10) that operates quantitatively at four different concentrations. Minimizing the number of tracers offers a significant workflow improvement over the previous protocol that utilized a combination of 6 tracers. Each NanoBRET(TM) kinase assay is built using commercially available plasmids and has been optimized for NanoLuc tagging orientation, diluent DNA, and tracer concentration. For complete details on the use and execution of this protocol, please refer to Vasta et al. (2018). Elsevier 2021-09-15 /pmc/articles/PMC8449129/ /pubmed/34568844 http://dx.doi.org/10.1016/j.xpro.2021.100822 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
Robers, Matthew B.
Wilkinson, Jennifer M.
Vasta, James D.
Berger, Lena M.
Berger, Benedict-Tilman
Knapp, Stefan
Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET
title Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET
title_full Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET
title_fullStr Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET
title_full_unstemmed Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET
title_short Single tracer-based protocol for broad-spectrum kinase profiling in live cells with NanoBRET
title_sort single tracer-based protocol for broad-spectrum kinase profiling in live cells with nanobret
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449129/
https://www.ncbi.nlm.nih.gov/pubmed/34568844
http://dx.doi.org/10.1016/j.xpro.2021.100822
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