Chapter title |
Express γ-H2AX Immunocytochemical Detection of DNA Damage
|
---|---|
Chapter number | 10 |
Book title |
Fast Detection of DNA Damage
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7187-9_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7185-5, 978-1-4939-7187-9
|
Authors |
Nate Hopp, Jodi Hagen, Birte Aggeler, Alexander E. Kalyuzhny |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 8 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Unspecified | 2 | 25% |
Student > Ph. D. Student | 2 | 25% |
Student > Master | 2 | 25% |
Student > Postgraduate | 1 | 13% |
Researcher | 1 | 13% |
Other | 0 | 0% |
Readers by discipline | Count | As % |
---|---|---|
Unspecified | 2 | 25% |
Biochemistry, Genetics and Molecular Biology | 2 | 25% |
Nursing and Health Professions | 1 | 13% |
Agricultural and Biological Sciences | 1 | 13% |
Sports and Recreations | 1 | 13% |
Other | 0 | 0% |
Unknown | 1 | 13% |