Lipid peroxidation probe bdp 581 591 c11

Malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) activities were quantified by MDA kits (Cat# A003-1), GSH kits (Cat# A005), and SOD kits (Cat# A001-1) (JianCheng, Nanjing, China), respectively. The levels of LPO of HK-2 cells in different groups were evaluated by Lipid Peroxidation Probe BDP 581/591 C11 (Cat# L267, Dojindo Laboratories, Japan) according to the manufacturer’s instructions. The assessment of reactive oxygen species (ROS) production was conducted using DCFH-DA and DHE staining, in accordance with the guidelines provided by the manufacturer, and was subsequently measured through microfluorimetry detection.

We used Lipid Peroxidation Probe -BDP 581/591 C11 for lipid peroxidation detection(Dojindo Laboratories, L267). The protocol we used was in accordance with the manufacturer's instructions, and involved following the specific steps outlined below: HL60 and MOLM13 cells were plated in a 12-well plate at a density of 6*10^5 cells/ml, with RSL3 at a concentration of 0.5 μM. After incubating in a cell culture incubator for 24 h, the culture medium was discarded. The cells were then washed twice with HBSS by centrifuging at 2000 rpm for 3 min to collect them. After removing the HBSS, probe working solution was added and the cells were incubated in the cell culture incubator for 30 min. The working solution was subsequently removed, and the cells were rinsed twice with HBSS before adding 500 µl of HBSS per well. Finally, the cells were observed and photographed using a fluorescence microscope.

FerroOrange (#F374), ROS Assay Kit -Photo-oxidation Resistant DCFH-DA (#R253) and Lipid Peroxidation Probe-BDP 581/591 C11 (#L267) were purchased from DOJINDO LABORATORIES (Kumamoto, Japan). Antibodies against GPX4 (#A1933), NCOA4 (#A5695), ACTB (#AC026), FTH1 (#A19544), LC3 (#A17424), KRAS (#A12704) as well as secondary HRP-conjugated Goat Anti-Rabbit IgG (#AS014), secondary HRP-conjugated Goat Anti-Mouse IgG (#AS003) were from ABclonal (Wuhan, China). Antibodies against CH25H (#ER1919-26) were from HUABIO (Hangzhou, China). Erastin (#S7242) was purchased from Selleckchem (Shanghai, China). Chloroquine (#HY-17589A) and wortmannin (#HY-10197) were purchased from MCE (Shanghai, China). 25-HC (#H1015) was purchased from Sigma-Aldrich (MO, USA).