Cargando…

The γH2AX DNA damage assay from a drop of blood

DNA double-strand breaks (DSB) and blocked replication forks activate the DNA damage response (DDR), a signaling pathway marked by phosphorylation of histone 2AX (H2AX). The phosphorylated form, γH2AX, accumulates at the site of damage and can be detected as foci by immunocytochemistry. Therefore, γ...

Descripción completa

Detalles Bibliográficos
Autores principales: Heylmann, Daniel, Kaina, Bernd
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4778029/
https://www.ncbi.nlm.nih.gov/pubmed/26940638
http://dx.doi.org/10.1038/srep22682
_version_ 1782419388157132800
author Heylmann, Daniel
Kaina, Bernd
author_facet Heylmann, Daniel
Kaina, Bernd
author_sort Heylmann, Daniel
collection PubMed
description DNA double-strand breaks (DSB) and blocked replication forks activate the DNA damage response (DDR), a signaling pathway marked by phosphorylation of histone 2AX (H2AX). The phosphorylated form, γH2AX, accumulates at the site of damage and can be detected as foci by immunocytochemistry. Therefore, γH2AX is a sensitive and robust biomarker of DNA damage, notably DSB. Cells from peripheral blood are often used for studies on genotoxic exposure of humans. They are limited, however, by the amount of blood required and the costly blood purification method. Here, we present a method that enables the detection of DNA damage by the analysis of γH2AX foci in a drop of blood. The blood drop method (BDM) is simple, fast, inexpensive and allows large series of blood sampling and storage over time. It can be combined with genotoxic treatment of cells in the collected blood sample for experimental purposes on DNA damage induction and repair. The BDM is suitable for rapid and large-scale screenings of genetic damage in human and animal populations.
format Online
Article
Text
id pubmed-4778029
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-47780292016-03-09 The γH2AX DNA damage assay from a drop of blood Heylmann, Daniel Kaina, Bernd Sci Rep Article DNA double-strand breaks (DSB) and blocked replication forks activate the DNA damage response (DDR), a signaling pathway marked by phosphorylation of histone 2AX (H2AX). The phosphorylated form, γH2AX, accumulates at the site of damage and can be detected as foci by immunocytochemistry. Therefore, γH2AX is a sensitive and robust biomarker of DNA damage, notably DSB. Cells from peripheral blood are often used for studies on genotoxic exposure of humans. They are limited, however, by the amount of blood required and the costly blood purification method. Here, we present a method that enables the detection of DNA damage by the analysis of γH2AX foci in a drop of blood. The blood drop method (BDM) is simple, fast, inexpensive and allows large series of blood sampling and storage over time. It can be combined with genotoxic treatment of cells in the collected blood sample for experimental purposes on DNA damage induction and repair. The BDM is suitable for rapid and large-scale screenings of genetic damage in human and animal populations. Nature Publishing Group 2016-03-04 /pmc/articles/PMC4778029/ /pubmed/26940638 http://dx.doi.org/10.1038/srep22682 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Heylmann, Daniel
Kaina, Bernd
The γH2AX DNA damage assay from a drop of blood
title The γH2AX DNA damage assay from a drop of blood
title_full The γH2AX DNA damage assay from a drop of blood
title_fullStr The γH2AX DNA damage assay from a drop of blood
title_full_unstemmed The γH2AX DNA damage assay from a drop of blood
title_short The γH2AX DNA damage assay from a drop of blood
title_sort γh2ax dna damage assay from a drop of blood
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4778029/
https://www.ncbi.nlm.nih.gov/pubmed/26940638
http://dx.doi.org/10.1038/srep22682
work_keys_str_mv AT heylmanndaniel thegh2axdnadamageassayfromadropofblood
AT kainabernd thegh2axdnadamageassayfromadropofblood
AT heylmanndaniel gh2axdnadamageassayfromadropofblood
AT kainabernd gh2axdnadamageassayfromadropofblood