Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 160, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/60682
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Funding
- National Institutes of Health [K01DK114390]
- American Cancer Society [RSG-18-050-01-NEC]
- University of New Mexico Environmental Health Signature Program and Superfund [P42 ES025589]
- UNM comprehensive cancer center [P30CA118100]
- Dedicated Health Research Funds at the University of New Mexico School of Medicine
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Oxidative stress is an important event under both physiological and pathological conditions. In this study, we demonstrate how to quantify oxidative stress by measuring total reactive oxygen species (ROS) using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining in colorectal cancer cell lines as an example. This protocol describes detailed steps including preparation of DCFH-DA solution, incubation of cells with DCFH-DA solution, and measurement of normalized intensity. DCFH-DA staining is a simple and cost-effective way to detect ROS in cells. It can be used to measure ROS generation after chemical treatment or genetic modifications. Therefore, it is useful for determining cellular oxidative stress upon environment stress, providing clues to mechanistic studies.
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