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

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.