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Open Access 01-12-2009 | Research

High throughput evaluation of gamma-H2AX

Authors: Dane Avondoglio, Tamalee Scott, Whoon Jong Kil, Mary Sproull, Philip J Tofilon, Kevin Camphausen

Published in: Radiation Oncology | Issue 1/2009

Abstract

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 γ-H2AX is a known marker for irradiation-induced DNA DSBs. However, the first generation assay involves the use of immunofluorescent staining of γ-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 γ-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 γ-H2AX versus irradiation dose.

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Title: High throughput evaluation of gamma-H2AX
Authors: Dane Avondoglio
Tamalee Scott
Whoon Jong Kil
Mary Sproull
Philip J Tofilon
Kevin Camphausen
Publication date: 01-12-2009
Publisher: BioMed Central
Published in: Radiation Oncology / Issue 1/2009
Electronic ISSN: 1748-717X
DOI: https://doi.org/10.1186/1748-717X-4-31

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High throughput evaluation of gamma-H2AX

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