Ferroptosis

Ferroptosis is a form of programmed cell death that is caused by iron ion-dependent accumulation of lipid peroxides. Ferroptosis has been shown to follow a different cell death pathway from apoptosis and thus is attracting attention as a new target for cancer therapy. It has also been found to be associated with various diseases, such as neurodegenerative diseases, cerebral apoplexy, and hepatitis (NASH).

Mechanism of Ferroptosis

Ferroptosis is characterized by the accumulation of lipid peroxides. Lipid peroxides are formed from oxidation of polyunsaturated fatty acids (PUFA) in membrane phospholipids, with iron suggested to be involved. Intracellular glutathione peroxidase 4 (GPX4) uses reduced glutathione (GSH), an antioxidant, to reduce lipid peroxides generated by reactive oxygen species (ROS).

However, when lipid peroxides accumulate due to GPX4 disruption or GSH depletion, ferroptosis is triggered.

Mechanism of Ferroptosis

Induction of Ferroptosis by Erastin

Erastin is a known inducer of ferroptosis. By inhibiting the cystine transporter (xCT), erastin inhibits the uptake of cystine. Cystine is the raw material for GSH. Therefore, Erastin ultimately decreases the amount of GSH. Decreased GSH then results in lipid peroxide accumulation and induction of ferroptosis.

The following experimental examples show changes in each index because of erastin stimulation.

Using erastin-treated A549 cells, we measured intracellular Fe2+, ROS, lipid peroxide, glutathione, glutamate release into the extracellular space, and cystine uptake. As a result, inhibition of xCT by elastin was observed and the release of glutamate and uptake of cystine were decreased. Furthermore, elastin treatment decreased intracellular glutathione while it increased intracellular Fe2+, ROS, and lipid peroxides.

Induction of Ferroptosis by Erastin

Object Product Name CAT. No.
①Cystine Uptake (Under Development)  
②Released Glutamate Glutamate Assay Kit-WST G269-10
③Intracellular Glutathione GSSG/GSH Quantification Kit G257-10
④Intracellular Fe2+ FerroOrange F374-10; F374-12
⑤Intracellular ROS ROS Assay Kit -Highly Sensitive DCFH-DA- R252-10
⑥Intracellular Lipid Peroxide Liperfluo L248-10

Ferroptosis-related Reagents Selection

Lipid Peroxide and Iron (Fe2+) Detection Reagents

Product Name CAT. No. Target Localization Detection (Fluorescence: Ex/Em) Instrument Sample
Liperfluo L248-10 Lipid Peroxidation Intracellular Fluorescence (524 nm/535 nm) Fluorescence Microscope, FCM Live Cell
MitoPeDPP M466-10 Lipid Peroxidation Mitochondria Fluorescence (452 nm/470 nm) Fluorescence Microscope, FCM Live Cell
Mito-FerroGreen M489-10 Ferrous Ion (Fe2+) Mitochondria Fluorescence (505 nm/580 nm) Fluorescence Microscope, Microplate Reader Live Cell
FerroOrange F374-10; F374-12 Ferrous Ion (Fe2+) Intracellular Fluorescence (543 nm/580 nm) Fluorescence Microscope Live Cell

Oxidative Stress- and Metabolism-Related Reagents and Kits

Name CAT. No. Target Localization Detection (Fluorescence: Ex/Em) Instrument Sample
ROS Assay Kit -Highly Sensitive DCFH-DA- R252-10 ROS (Reactive oxygen species) Intracellular Fluorescence
(505 nm/525 nm)
Fluorescence Microscope, FCM, Microplate Reader Live Cell
GSSG/GSH Quantification Kit G257-10 Glutathione (oxidized/reduced) Intracellular Colorimetric:412 nm Microplate Reader Cell, Tissue, Blood Plasma, Red Blood Cell
Glutamine Assay Kit-WST G268-10 Glutamine Intracellular/ Extracellular Colorimetric:450 nm Microplate Reader Cell, Cell Culture
Glutamate Assay Kit-WST G269-10 Glutamine Intracellular/ Extracellular Colorimetric:450 nm Microplate Reader Cell, Cell Culture

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