Express γ-H2AX Immunocytochemical Detection of DNA Damage.
Methods Mol Biol
; 1644: 123-128, 2017.
Article
em En
| MEDLINE
| ID: mdl-28710758
ABSTRACT
DNA can be damaged by many environmental factors including chemical agents and ionizing radiation which induce the formation of DNA double-stranded breaks (DSBs). If DSBs are not repaired in a timely fashion this may cause the disruption of genome integrity, which can result in cancer development. Typically, DSBs are followed by phosphorylation of histone protein H2AX, a member of the H2A family. Immunocytochemical detection of phosphorylated H2AX (e.g., γ-H2AX) appears to be a useful technique for assessing DNA damage. Such an assessment is easy to do by analyzing labeling for γ-H2AX under the microscope and does not require an expensive laboratory setup. Using HeLa cells treated with camptothecin as a model, we developed an easy-to-run protocol to analyze DSBs. Our protocol can be applied to testing the potency of different chemicals to induce DSBs in different types of cells and requires around 2 h to complete.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Camptotecina
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Imuno-Histoquímica
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Histonas
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Reparo do DNA
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Quebras de DNA de Cadeia Dupla
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article