Elevated iron level and accumulated lipid ROS were representative characteristics of ferroptosis. We used an iron assay kit (Beyotime) to examine the level of intracellular Fe2+. Briefly, the cells were homogenized to collect the supernatant, incubated with iron reducer, followed by labeling by iron probe. The OD 590 nm were detected in a microplate reader (PerkinElmer). For detection of lipid ROS, cells were stained with BODIPY C-11 dye (Beyotime) for 30 min, and subsequently detected by Flow cytometry (BD bioscience).
Bodipy c 11 dye
Manufactured by Beyotime
Sourced in United States
BODIPY C-11 dye is a fluorescent probe that can be used for various biological applications. It has an excitation wavelength of 522 nm and an emission wavelength of 544 nm. BODIPY C-11 dye is useful for detecting and tracking lipid-based molecules within cells.
Automatically generated - may contain errors
2 protocols using bodipy c 11 dye
1
Ferroptosis Characterization Protocol
2
Ferroptosis Induction and Measurement
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SGC7901 and BGC823 cells were cotreated with erastin (5 mmol/L) or ferrostatin (1 mmol/L). After 48 h, the cell viability was analyzed by MTT assay. Elevated iron level and accumulated lipid reactive oxygen species (ROS) were representative characteristics of ferroptosis. We used an iron assay kit (Beyotime, China) to examine the level of intracellular Fe2+. The cells were homogenized to collect the supernatant, incubated with iron reducer, followed by labeling with iron probe. OD 590 nm was detected in a microplate reader (PerkinElmer, Waltham, MA, United States). For detection of lipid ROS, cells were stained with BODIPY C-11 dye (Beyotime) for 30 min, and subsequently detected by flow cytometry (BD Biosciences, Franklin Lakes, NJ, United States). The levels of malondialdehyde (MDA) and glutathione peroxidase (GSH) was measured by the MDA detection kit (Beyotime) and GSH assay kit (Cayman, Ann Arbor, MI, United States), respectively.
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