High throughput evaluation of gamma-H2AX

被引：33

作者：

Avondoglio, Dane ^{[1
]}

Scott, Tamalee ^{[1
]}

Kil, Whoon Jong

Sproull, Mary ^{[1
]}

Tofilon, Philip J. ^{[2
]}

Camphausen, Kevin ^{[1
]}

机构：

[1] NCI, Radiat Oncol Branch, Bethesda, MD 20892 USA

[2] Univ S Florida, H Lee Moffitt Canc Ctr, Drug Discovery Program, Tampa, FL 33682 USA

来源：

RADIATION ONCOLOGY | 2009年 / 4卷

关键词：

STRAND BREAK REPAIR; HISTONE H2AX; RADIOSENSITIZATION; INHIBITION; EXPRESSION; CELLS;

D O I：

10.1186/1748-717X-4-31

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

The DNA double-strand break (DSB) is the primary lethal lesion after therapeutic radiation. Thus, the development of assays to detect and to quantitate these lesions could have broad preclinical and clinical impact. Phosphorylation of histone H2AX to form gamma-H2AX is a known marker for irradiation-induced DNA DSBs. However, the first generation assay involves the use of immunofluorescent staining of gamma-H2AX foci. This assay is time consuming, operator dependent and is not scalable for high throughput assay development. Thus, we sought to develop a new assay using a high throughput electrochemiluminescent platform from Mesoscale Discovery Systems to quantify gamma-H2AX levels. The results show that our assay utilizes significantly less time and labor, has greater intra-assay reproducibility and has a greater dynamic range of gamma-H2AX versus irradiation dose.

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页数：5

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