Quantifying DNA Double-Strand Breaks via γH2AX

Since γH2AX is a protein for targeting DNA double-strand breaks, monitoring the relationship of γH2AX and cellular DNA is reasonable. For the γH2AX assay, p-Histone H2A.X antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used to recognize the phosphorylated H2AX (γH2AX). Then, the Alexa Fluor^®488-secondary antibody (Cell Signaling Technology, Danvers, MA, USA) was applied [62 (link)]. 7AAD (1 μg/mL, 30 min) was further applied to stain cellular DNA. Both γH2AX and 7AAD (+) populations were examined for DNA damage with double-strand breaks. The Accuri C6 flow cytometer detected the Alexa Fluor^®488 intensities for γH2AX contents.

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Yang K.H., Lin Y.S., Wang S.C., Lee M.Y., Tang J.Y., Chang F.R., Chuang Y.T., Sheu J.H, & Chang H.W. (2021). Soft Coral-Derived Dihydrosinularin Exhibits Antiproliferative Effects Associated with Apoptosis and DNA Damage in Oral Cancer Cells. Pharmaceuticals, 14(10), 994.

Publication 2021

p-Histone H2A.X antibody Alexa Fluor®488-secondary antibody

7aad Alexa fluor 488 Antibody Assay Cellular Dna damage Dna double strand breaks Histone h2a x Populations Stain

Corresponding Organization : China Medical University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Phosphorylated H2AX (γH2AX) content
7AAD (+) population (cellular DNA)

control variables

None explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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