High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation

DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn’t accumulate in...

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Autores principales: Oeck, Sebastian, Malewicz, Nathalie M., Krysztofiak, Adam, Turchick, Audrey, Jendrossek, Verena, Glazer, Peter M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6395640/
https://www.ncbi.nlm.nih.gov/pubmed/30816253
http://dx.doi.org/10.1038/s41598-019-39760-8
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author Oeck, Sebastian
Malewicz, Nathalie M.
Krysztofiak, Adam
Turchick, Audrey
Jendrossek, Verena
Glazer, Peter M.
author_facet Oeck, Sebastian
Malewicz, Nathalie M.
Krysztofiak, Adam
Turchick, Audrey
Jendrossek, Verena
Glazer, Peter M.
author_sort Oeck, Sebastian
collection PubMed
description DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn’t accumulate in foci, laser micro-irradiation of single nuclei provides an alternative method to monitor DNA repair proteins and histone dynamics at the DNA damage site. We have developed an automated evaluation tool for standardized, high-throughput analysis of micro-irradiated cells featuring single cell background subtraction and detection across multiple fluorescence channels, allowing for robust statistics.
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spelling pubmed-63956402019-03-04 High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation Oeck, Sebastian Malewicz, Nathalie M. Krysztofiak, Adam Turchick, Audrey Jendrossek, Verena Glazer, Peter M. Sci Rep Article DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn’t accumulate in foci, laser micro-irradiation of single nuclei provides an alternative method to monitor DNA repair proteins and histone dynamics at the DNA damage site. We have developed an automated evaluation tool for standardized, high-throughput analysis of micro-irradiated cells featuring single cell background subtraction and detection across multiple fluorescence channels, allowing for robust statistics. Nature Publishing Group UK 2019-02-28 /pmc/articles/PMC6395640/ /pubmed/30816253 http://dx.doi.org/10.1038/s41598-019-39760-8 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Oeck, Sebastian
Malewicz, Nathalie M.
Krysztofiak, Adam
Turchick, Audrey
Jendrossek, Verena
Glazer, Peter M.
High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation
title High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation
title_full High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation
title_fullStr High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation
title_full_unstemmed High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation
title_short High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation
title_sort high-throughput evaluation of protein migration and localization after laser micro-irradiation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6395640/
https://www.ncbi.nlm.nih.gov/pubmed/30816253
http://dx.doi.org/10.1038/s41598-019-39760-8
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