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Protocol to evaluate markers of reactive oxygen species and labile iron from a single sample across in vitro and in in vivo lung cancer models

Year: 2026

Type: article

Abstract: Cancer cells have dysregulated oxidative metabolism, elevated levels of reactive oxygen species (ROS), and a dependency on redox-active iron that supports tumorigenesis. We present a protocol for evaluating the intracellular oxidative environment and labile iron in cell culture and mouse models of lung cancer. We describe steps for harvesting samples, fluorescent antibody staining, staining for ROS and iron markers, and spectral flow cytometry analysis. This protocol allows insight into the role of redox stress and redox-active metals in the tumor microenvironment preclinically.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104596

Authors: Jennifer A. Petsche, Sunny C. Huang, Danielle J. Foster, Charles C. Searby, Val C. Sheffield, Douglas R. Spitz, Calvin S. Carter

Institutions: University of Iowa Stead Family Children’s Hospital, University of Iowa Health Care, University of Iowa Holden Comprehensive Cancer Center, University of Iowa Hospitals and Clinics

Language: English

Topic: Ferroptosis and cancer prognosis

Subfield: Pulmonary and Respiratory Medicine

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550

Funders: National Institutes of Health, Holden Comprehensive Cancer Center, University of Iowa, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, National Cancer Institute

Awards: Evaluation of Orthogonally Oriented Electromagnetic Fields to Stabilize ROS, Induce DNA damage and Improve Survival in Non-Small Cell Lung Cancer, Training Program in Free Radical and Radiation Biology, Cancer Center Support Grant

OpenAlex ID: W7163955566

FWCI:

Cites: 20

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Related to: 10