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Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers

The efficacy of cellular signal transduction is of paramount importance for the proper functioning of a cell and an organism as a whole. Protein kinases are responsible for much of this transmission and thus have been the focal point of extensive research. While there are numerous commercially avail...

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Detalles Bibliográficos
Autores principales: Bell, Ryan A.V., Storey, Kenneth B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4472853/
https://www.ncbi.nlm.nih.gov/pubmed/26150941
http://dx.doi.org/10.1016/j.mex.2014.07.003
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author Bell, Ryan A.V.
Storey, Kenneth B.
author_facet Bell, Ryan A.V.
Storey, Kenneth B.
author_sort Bell, Ryan A.V.
collection PubMed
description The efficacy of cellular signal transduction is of paramount importance for the proper functioning of a cell and an organism as a whole. Protein kinases are responsible for much of this transmission and thus have been the focal point of extensive research. While there are numerous commercially available protein kinase assays, the Kinase-Glo luminescent kinase assay (Promega) provides an easy-to-use and high throughput platform for determining protein kinase activity. This assay is said to require the use of a microplate spectrophotometer capable of detecting a luminescent signal. This study shows that: • The ChemiGenius Bioimaging system (Syngene), typically used for visualizing chemiluminescence from Western blots, provides an alternative detection system for Kinase-Glo luminescence. • The novel detection system confers an advantage over traditional luminometers, in that it allows visualization of the luminescent wells, which allows for the real-time analysis and correction of experimental errors (i.e. bubble formation). • Determining kinase kinetics using this detection system produced comparable results to previous studies on the same enzyme (i.e. glycogen synthase kinase 3).
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spelling pubmed-44728532015-07-06 Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers Bell, Ryan A.V. Storey, Kenneth B. MethodsX Article The efficacy of cellular signal transduction is of paramount importance for the proper functioning of a cell and an organism as a whole. Protein kinases are responsible for much of this transmission and thus have been the focal point of extensive research. While there are numerous commercially available protein kinase assays, the Kinase-Glo luminescent kinase assay (Promega) provides an easy-to-use and high throughput platform for determining protein kinase activity. This assay is said to require the use of a microplate spectrophotometer capable of detecting a luminescent signal. This study shows that: • The ChemiGenius Bioimaging system (Syngene), typically used for visualizing chemiluminescence from Western blots, provides an alternative detection system for Kinase-Glo luminescence. • The novel detection system confers an advantage over traditional luminometers, in that it allows visualization of the luminescent wells, which allows for the real-time analysis and correction of experimental errors (i.e. bubble formation). • Determining kinase kinetics using this detection system produced comparable results to previous studies on the same enzyme (i.e. glycogen synthase kinase 3). Elsevier 2014-08-08 /pmc/articles/PMC4472853/ /pubmed/26150941 http://dx.doi.org/10.1016/j.mex.2014.07.003 Text en © 2014 The Authors http://creativecommons.org/licenses/by/3.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Bell, Ryan A.V.
Storey, Kenneth B.
Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers
title Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers
title_full Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers
title_fullStr Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers
title_full_unstemmed Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers
title_short Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers
title_sort novel detection method for chemiluminescence derived from the kinase-glo luminescent kinase assay platform: advantages over traditional microplate luminometers
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4472853/
https://www.ncbi.nlm.nih.gov/pubmed/26150941
http://dx.doi.org/10.1016/j.mex.2014.07.003
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