Iron assay kit

Manufactured by Dojindo Laboratories

Sourced in Japan

The Iron Assay Kit is a laboratory product designed to quantify the amount of iron present in a sample. It provides a simple and reliable method for determining iron concentration through a colorimetric reaction. The kit includes all the necessary reagents and buffers to perform the analysis.

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Lab products found in correlation

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22 protocols using iron assay kit

Quantifying Tumor Iron Levels

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The Fe²⁺ level in vivo within the tumor was evaluated by Iron assay kit. Briefly, when the tumor size reached approximately 100–150 mm³, FGP (GOx 300 μg/mL, Fe 500 μg/mL) and saline were intravenously injected into 4T1 tumor-bearing mice (n = 3). Thereafter, all mice were sacrificed and the tumors were collected after 24 h post-injection. To measure the Fe²⁺ content, the tumors were weighed and homogenized in 1 mL of ice-cold PBS by a homogenizer and centrifuged (16,000×g) (Foring technology) for 10 min at 4 °C. Then, the level of Fe²⁺ in the supernatants were determined by the corresponding Iron Assay Kits (Dojindo, Tokyo, Japan) according to the manufacture's instructions.

Ren H., Yong J., Yang Q., Yang Z., Liu Z., Xu Y., Wang H., Jiang X., Miao W, & Li X. (2021). Self-assembled FeS-based cascade bioreactor with enhanced tumor penetration and synergistic treatments to trigger robust cancer immunotherapy. Acta Pharmaceutica Sinica. B, 11(10), 3244-3261.

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Quantifying Fe2+ and Total Iron

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To measure the Fe 2+ and total iron content, the cardiac tissues were weighed, homogenized and centrifuged (16,000g) for 10 min. The supernatant for quantitative determination of Fe 2+ and total iron content was collected, then measured using Iron Assay Kits (I291, Dojindo) according to manufacturer's instruction.

Zhang L.L., Chen G.H., Tang R.J., Xiong Y.Y., Pan Q., Jiang W.Y., Gong Z.T., Chen C., Li X.S, & Yang Y.J. (2024). Levosimendan Reverses Cardiac Malfunction and Cardiomyocyte Ferroptosis During Heart Failure with Preserved Ejection Fraction via Connexin 43 Signaling Activation. Cardiovascular drugs and therapy, 38(4).

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Measuring Endogenous Lung Iron Levels

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The concentrations of endogenous ferrous iron levels of the lung tissue were measured with the iron assay kit (I291, Dojindo, Japan). Lung tissue was fully washed with cold PBS, and homogenized in iron assay buffer on the ice, and then centrifuged at 16,000 × g for 10 min. The supernatant was collected for subsequent experiments. The 100 μL standard (1 mmol/L) was added with 900 μL assay buffer to made into 1000 μmol/l standard solution and then diluted to prepare standard solution (50, 25, 12.5, 6.25, 3.125, 1.5625, 0 μmol/l). The 5% volume of reducer solution was added to each of standard solution, and 5% volume of assay buffer for ferrous iron along with blank tube. All tubes were incubated for 15 min at 37 °C. The 100 μL iron probe was added to each reaction, then mixed and incubated for a further 60 min at 37 °C in a 96-well plate. The absorbance at 593 nm was measured by microplate reader to calculate the iron level.

Guo B., Zuo Z., Di X., Huang Y., Gong G., Xu B., Wang L., Zhang X., Liang Z., Hou Y., Liu X, & Hu Z. (2022). Salidroside attenuates HALI via IL-17A-mediated ferroptosis of alveolar epithelial cells by regulating Act1-TRAF6-p38 MAPK pathway. Cell Communication and Signaling : CCS, 20, 183.

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Quantifying Lung Iron Levels

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The levels of iron in lung tissue were measured by Iron Assay Kit (I291, Dojindo, Japan) according to the manufacturer’s instructions.

Ma L., He Y., Xie H., Wang J., Chen J., Song S., Zhang L., Li L., Lai H., Liu Y., Chen H., Zhang X., Liu X., Zou Z., Zhang Q., Yan J, & Tao A. (2024). Ferroptotic alveolar epithelial type II cells drive TH2 and TH17 mixed asthma triggered by birch pollen allergen Bet v 1. Cell Death Discovery, 10, 96.

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Quantitative Analysis of Oxidative Stress

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The cartilage tissues and chondrocytes were lysed using Cell lysis buffer (Solarbio, China) and detected with BCA kit (Servicebio, China). The MDA and GSH concentration were detected using MDA Assay Kit (Beyotime, China) or GSH/GSSG Assay Kit (Beyotime, China) and normalized based on the protein concentration. The Fe²⁺ detection was performed using Iron Assay Kit (Dojindo, Japan) according to the kit’s instructions.

Kong R., Ji L., Pang Y., Zhao D, & Gao J. (2023). Exosomes from osteoarthritic fibroblast-like synoviocytes promote cartilage ferroptosis and damage via delivering microRNA-19b-3p to target SLC7A11 in osteoarthritis. Frontiers in Immunology, 14, 1181156.

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Intracellular Iron Quantification

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Intracellular ferrous ion was determined using Iron Assay Kit (I291, Dojindo) following the manufacturer’s instructions. The values were measured using a microplate reader at the absorption wavelength of 593 nm. The iron concentration in tumor tissues was assessed using the Prussian Blue Iron Stain Kit (G1428, Solarbio Life Sciences) according to the manufacturer’s instructions. Images were obtained using an Olympus CX31 microscope.

Kang L., Wang D., Shen T., Liu X., Dai B., Zhou D., Shen H., Gong J., Li G., Hu Y., Wang P., Mi X., Zhang Y, & Tan X. (2023). PDIA4 confers resistance to ferroptosis via induction of ATF4/SLC7A11 in renal cell carcinoma. Cell Death & Disease, 14(3), 193.

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Cellular and Mitochondrial Iron Quantification

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Cellular and mitochondrial ferrous iron concentrations were assessed using the FerroOrange probe (Dojindo, Japan) and Mito-FerroGreen probe (Dojindo), respectively. Briefly, 5 × 10⁵ cells/well were seeded in 6-well plates and treated with 1 μmol/L SHK with or without DFOM. After 24 h, 1 μmol/L FerroOrange and 2 μmol/L Mito-FerroGreen probes were added to the cell culture, and the plates were incubated at 37°C for 30 min. The stained cells were monitored using a BD Accuri C6 flow cytometer (BD Biosciences). The tissue ferrous iron and total iron concentrations were assessed using an iron assay kit (Dojindo) according to the manufacturer’s instructions.

Li W., Fu H., Fang L., Chai H., Gao T., Chen Z, & Qian S. (2022). Shikonin induces ferroptosis in multiple myeloma via GOT1-mediated ferritinophagy. Frontiers in Oncology, 12, 1025067.

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Quantifying Iron Levels in OA Cartilage

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Human OA cartilage Iron ion detection were performed using Iron Assay Kit (Dojindo, Kumamoto, Japan) according to the manufacturer's instructions. Briefly, 100mg fresh human OA cartilage tissue was placed in a sterile 2ml tube containing 1 ml assay buffer, and were then grinded using homogenizer on the ice at 4°C. Then the homogenate was centrifuged at 16,000 g for 10 min, and the supernatant was collected. Assay buffer was added in supernatant to test Fe²⁺ level, or reducer solution was added to test total Iron level. The absorbance of the wells at a wavelength of 593 nm was measured on a microplate reader after a 1 h incubation at 37°C. Then the Fe²⁺, Fe³⁺ and total Iron levels of OA cartilage and corresponding non-lesion cartilage were calculated.

Miao Y., Chen Y., Xue F., Liu K., Zhu B., Gao J., Yin J., Zhang C, & Li G. (2022). Contribution of ferroptosis and GPX4’s dual functions to osteoarthritis progression. EBioMedicine, 76, 103847.

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Quantification of Oxidative Stress Markers

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The MDA levels in plasma and heart tissues were determined using the Colorimetric MDA Assay Kit (Jiancheng, China), and the Fe²⁺ and Fe³⁺ levels in heart tissues were determined by Iron Assay Kit (DOJINDO, Japan) per manufacturers’ instructions.

Pan J., Xiong W., Zhang A., Zhang H., Lin H., Gao L., Ke J., Huang S., Zhang J., Gu J., Chang A.C, & Wang C. (2023). The Imbalance of p53–Park7 Signaling Axis Induces Iron Homeostasis Dysfunction in Doxorubicin‐Challenged Cardiomyocytes. Advanced Science, 10(15), 2206007.

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Quantifying Mouse Testis Iron Levels

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The iron content level in the mouse testes was evaluated using an iron assay kit (Dojindo, Tokyo, Japan). Briefly, fresh mouse testes were homogenized in PBS. After centrifugation, the supernatant was collected, and total Fe, Fe²⁺, and Fe³⁺ levels were measured using an iron analysis kit (Dojindo, Tokyo, Japan) according to the manufacturer's instructions.

Zhao Y., Zhang H., Cui J.G., Wang J.X., Chen M.S., Wang H.R., Li X.N, & Li J.L. (2022). Ferroptosis is critical for phthalates driving the blood-testis barrier dysfunction via targeting transferrin receptor. Redox Biology, 59, 102584.

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