Iron content in ESCC cells was monitored with iron assay kit in line with the manufacturer's instruction (Beyotime, China). Cells were homogenized via five volumes of iron assay buffer and centrifugated at 13,000 g lasting 10 min at 4°C. Thereafter, iron reducer was added to supernatant mixture and treated for 30 min at room temperature, followed by iron probe protecting against light for 1 h. The absorbance value was monitored at 593 nm.
Iron assay kit
Manufactured by Beyotime
Sourced in China
The Iron Assay Kit is a quantitative colorimetric assay designed to measure the concentration of iron in a variety of sample types. The kit utilizes a chromogenic reagent that reacts with iron to produce a colored complex, which is then measured spectrophotometrically.
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Lab products found in correlation
5 protocols using iron assay kit
1
Quantifying Iron in ESCC Cells
2
Ferroptosis Characterization Protocol
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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).
3
Measuring Iron Levels in H/R-Induced HMCs
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The levels of iron ion in H/R-induced HMCs were measured using iron assay kit (Beyotime, China) following the manufacturer's instructions. Briefly, in the 5-10 X volumes of iron assay buffer, H/R-induced HMCs were homogenized. Cell supernatant were then added with iron reducer and incubated for 20 min, and the absorbance at 593 nm of each group was assessed by the microplate reader.
4
Intracellular Iron and Lipid ROS Assay
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Iron assay kit (Beyotime) and BODIPY C-11 (Sigma) staining were conducted to measure the intracellular Fe2+ level and the lipid ROS level in SKOV-3 and T47D cells under the manufacturers’ instructions, separately.
5
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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