Journal article
Protocol to evaluate markers of reactiveoxygen species and labile iron from a single sample across in vitro and in in vivo lung cancer models
STAR protocols, Vol.7(2), 104596
06/08/2026
DOI: 10.1016/j.xpro.2026.104596
PMCID: PMC13264167
PMID: 42258355
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.
[Display omitted]
•Instructions for harvesting and plating samples for flow cytometry analysis•Guidance on fluorescent antibody staining•Steps for evaluating markers of reactive oxygen species (ROS) and labile iron
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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.
Details
- Title: Subtitle
- Protocol to evaluate markers of reactiveoxygen species and labile iron from a single sample across in vitro and in in vivo lung cancer models
- Creators
- Jennifer A. Petsche - University of IowaSunny C. Huang - University of IowaDanielle J. Foster - University of IowaCharles C. Searby - University of IowaVal C. Sheffield - University of IowaDouglas R. Spitz - University of IowaCalvin S. Carter - University of Iowa
- Resource Type
- Journal article
- Publication Details
- STAR protocols, Vol.7(2), 104596
- DOI
- 10.1016/j.xpro.2026.104596
- PMID
- 42258355
- PMCID
- PMC13264167
- NLM abbreviation
- STAR Protoc
- ISSN
- 2666-1667
- eISSN
- 2666-1667
- Publisher
- Elsevier Inc
- Grant note
- Carver College of Medicine Holden Comprehensive Cancer Center Iowa City Veteran's Administration Medical Center National Cancer Institute of the National Institutes of Health: R21CA256301, T32CA078586, P30CA086862
We would like to thank members of the Carter, Sheffield, and Spitz laboratories; Heath Vignes; Ann Miller; and Kranti Mapuskar for valuable discussions. The data presented herein were obtained at the Flow Cytometry Facility, which is a Carver College of Medicine/Holden Comprehensive Cancer Center (HCCC) core research facility at the University of Iowa. The facility is funded through user fees and the generous financial support of the Carver College of Medicine, the Holden Comprehensive Cancer Center, and the Iowa City Veteran's Administration Medical Center. Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under grant nos. R21CA256301 (C.S.C., V.C.S., and D.R.S.), T32CA078586 (S.C.H.), and P30CA086862 (HCCC).
- Language
- English
- Date published
- 06/08/2026
- Academic Unit
- Stead Family Department of Pediatrics; Pathology; Iowa Neuroscience Institute; Medical Genetics and Genomics; Radiation Oncology; Fraternal Order of Eagles Diabetes Research Center; Ophthalmology and Visual Sciences
- Record Identifier
- 9985174501502771
Metrics
1 Record Views