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Probing Lipid Peroxidation in Ferroptosis: Emphasizing the Utilization of C11-BODIPY-Based Protocols

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Ferroptosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2712))

Abstract

Ferroptosis is a form of regulated cell death that relies on iron and is characterized by the accumulation of lipid peroxides, resulting in oncotic cell swelling and eventual disruption of cellular membranes. Lipid peroxidation, a hallmark of ferroptosis, refers to the oxidative deterioration of lipids that contain carbon-carbon double bonds, particularly polyunsaturated fatty acids (PUFAs). Understanding the molecular mechanisms underlying the interplay between ferroptosis and lipid peroxidation and identifying reliable techniques for assessing lipid peroxidation levels are crucial for further advancements in this field of research. Various methods have been developed to detect lipid peroxidation levels, including C11-BODIPY (BODIPY™ 581/591 C11), liperfluo, 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), Click-iT LAA (linoleamide alkyne), and liquid chromatography-mass spectrometry (LC-MS)-based epilipidomics (redox lipidomics). Currently, one of the most commonly used and effective methods is the C11-BODIPY assay, which utilizes a fluorescent probe that selectively sensitizes lipid peroxidation in cell membranes. Incorporating advanced techniques such as flow cytometry and fluorescence microscopy with C11-BODIPY dye is essential for accurate assessment of lipid peroxidation levels in ferroptosis. This chapter aims to provide comprehensive experimental protocols for detecting lipid peroxidation levels indicative of ferroptosis using C11-BODIPY staining and subsequent detection via flow cytometry and fluorescence microscopy.

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Acknowledgements

We would like to express our sincere gratitude to Ziqi Wang for her invaluable assistance in creating the schematic diagram, and to Dr. Huabin Wang for proofreading the manuscript. This work was supported by grants from the Guangzhou Basic and Applied Basic Research Foundation (grant number 202102020509), the Program for Guangdong Introducing Innovative and Entrepreneurial Teams (grant number 2017ZT07S347), and the National Natural Science Foundation of China (grant number 31701174).

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Author notes

  1. Authors Zhangshuai Dai and Wanting Zhang have equally contributed to this chapter.

Authors and Affiliations

  1. Key Laboratory for Regenerative Medicine, Ministry of Education, College of Life Science and Technology, Jinan University, Guangzhou, China

    Zhangshuai Dai, Wanting Zhang, Liqun Zhou & Junqi Huang

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  1. Zhangshuai Dai
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  2. Wanting Zhang
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  3. Liqun Zhou
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  4. Junqi Huang
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Editors and Affiliations

  1. Centre de Recherche des Cordeliers, Université de Paris, Paris, France

    Guido Kroemer

  2. Department of Surgery, University of Texas Southwestern Medical Center, Dallas, TX, USA

    Daolin Tang

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Dai, Z., Zhang, W., Zhou, L., Huang, J. (2023). Probing Lipid Peroxidation in Ferroptosis: Emphasizing the Utilization of C11-BODIPY-Based Protocols. In: Kroemer, G., Tang, D. (eds) Ferroptosis. Methods in Molecular Biology, vol 2712. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3433-2_6

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  • DOI: https://doi.org/10.1007/978-1-0716-3433-2_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3432-5

  • Online ISBN: 978-1-0716-3433-2

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