02 Oxidative Stress

Iron Assay Kit -Colorimetric-

Oxidative Stress

Tissue Iron Detection

Detect iron in tissue samples
Standard samples for quantifying the content of Fe²⁺ and Fe³⁺ are included
A convenient calculation sheet for calculating iron content is provided

Product code

I291　 Iron Assay Kit -Colorimetric-

Unit size	Price	Item Code
50 tests	Find your distributors	I291-10

Component

50 tests	Probe Solution Assay Buffer Standard Reducer	7 ml x 1 20 ml x 1 150 μl (1 mmol/l) x 1 x 1

C of A

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Description

Iron is one of the most important metallic elements in living organisms. In recent years, an iron-dependent mode of cell death, ferroptosis, has emerged, and research on intracellular iron ions has attracted increased attention.
The Iron Assay Kit-Colorimetric is a kit for measuring the amount of iron ions (Fe²⁺ and Fe³⁺) in tissue. It enables data acquisition in approximately 2 h from sample pretreatment to the measurement of iron ion concentration.
The concentration of Fe²⁺ in a tissue sample can be determined by adding Ferene S, which is a chromogenic dye for Fe²⁺, to the tissue lysate and measuring its absorbance. By using the reducing agent included in the kit to reduce Fe³⁺ in the sample to Fe²⁺, the concentrations of total iron and Fe³⁺ in the sample can also be determined.

Ferroptosis Analysis Products

Product Name	Target	Ditection Properties
FerroOrange	Intracellular Fe²⁺	Microscopy, Plate reader Ex: 543 nm / Em: 580 nm
Mito-FerroGreen	Mitochondrial Fe²⁺	Microscopy Ex: 505 nm / Em: 535 nm
Lyso-FerroRed	Lysosomal Fe²⁺	Microscopy, FCM, Plate reader Ex: 551 nm / Em: 571 nm
Iron Assay Kit -Colorimetric-	Fe²⁺ and Fe³⁺	Plate reader Colorimetric, λ: 593 nm
Liperfluo	Lipid Peroxide	Microscopy, FCM Ex: 488 nm / Em: 500-550 nm
Lipid Peroxidation Probe -BDP 581/591 C11-	Lipid Peroxidation Process	Microscopy, FCM, Plate reader Pre-reaction, Ex: 488 nm / Em: 510-550 nm Post-reaction, Ex: 561 nm / Em: 600-630 nm
MDA Assay Kit	Malondialdehyde	Plate reader Fluorescence, Ex: 540 nm / Em: 590 nm Colorimetric, λ: 532 nm
Cystine Uptake Assay Kit	Cystine uptake	Plate reader Ex: 485 nm / Em: 535 nm
GSSG/GSH Quantification Kit	GSSG and GSH	Plate reader Colorimetric, λ: 405 nm

Manual

Technical info

Iron Detection Kit and Staining Reagents

Select the appropriate reagent based on your experimental method and detecting equipment.

	Lyso-FerroRed	FerroOrange	Mito-FerroGreen	Iron Assay Kit -Colorimetric-
Intracellular localization	Lysosome	Intracellular	Mitochondria	ー (Tissue)
Detection properties	Fluorometric λex : 551 nm, λem : 571 nm	Fluorometric λex : 543 nm, λem : 580 nm	Fluorometric λex : 505 nm, λem : 535 nm	Colorimetric 593 nm
Supported devices （filter）	Microscope, Plate reader, Flow cytometry(TRITC)	Microscope, Plate reader(Cy3)	Microscope(FITC, GFP)	Plate reader
Target	Lysosomal Fe²⁺ in live cells	Intracellular Fe²⁺ in live cells	Mitochondrial Fe²⁺ in live cells	Fe²⁺ and Fe³⁺ in tissue
Usage count	35 nmol can be used for 17 assays at 35 mm dish (final concentration 1 µmol/l).	1 tube (24 µg) can be used for 17 assays at 35 mm dish (final concentration 1 µmol/L)	1 set (50 µg x 2) can be used for 10 assays at 35 mm dish (final concentration 5 µmol/L)	50 tests (Make standard curve and detect Fe²⁺ only: 4 samples, detect Fe²⁺ and Fe³⁺: 2 samples)

Ferrous Ion Selectivity

Experimental example: Detection of Fe²⁺ and Fe³⁺ in liver sample

Using this kit, we measured the amounts of Fe²⁺ and Fe³⁺ in mouse liver. We confirmed that 100 mg of mouse liver contained 2.51 nmol of Fe²⁺, 3.05 nmol of Fe³⁺, and a total iron content of 5.56 nmol.

References

Open References

No.	Sample	Reference
1	Mouse tumor	H. Ren, J. Yong, Q. Yang, Z. Liu, Y. Xu, H. Wang, X. Jiang, W. Miao, X. Li, “Self-assembled FeS -based cascade bioreactor with enhanced tumor penetration and synergistic treatments to trigger robust cancer immunotherapy”, Acta Pharm. Sin. B, 2021, doi:10.1016/j.apsb.2021.05.005.
2	Human cartilage	H. Ren, J. Yong, Q. Yang, Z. Yang, Z. Liu, Y. Xu, H. Wang, X. Jiang, W. Miao, X. Li, "Contribution of ferroptosis and GPX4’s dual functions to osteoarthritis progression", EBioMedicine, 2022, doi: 10.1016/j.ebiom.2022.103847.
3	Mouse lung, liver, and kidney	T. Wu, X. Wang, M. Chen, X. Zhang, J. Zhang, J. Cheng, L. Kong, M. Tang,"Respiratory exposure to graphene quantum dots causes fibrotic effects on lung, liver and kidney of mice", Food Chem. Toxicol., 2022, doi: 10.1016/j.fct.2022.112971.
4	Mouse hippocampus	T. Wu, X. Wang, J. Cheng, X. Liang, Y. Li, M. Chen, L. Kong and M. Tang, "Nitrogen‑doped graphene quantum dots induce ferroptosis through disrupting calcium homeostasis in microglia", Part. Fibre Toxicol., 2022, doi: 10.1186/s12989-022-00464-z.
5	Human cervical mucosa	T. Wang, M. Gong, Y. Cao, C. Zhao, Y. Lu, Y. Zhou, S. Yao, J. Chen, C. Zhao and R. Ju, "Persistent ferroptosis promotes cervical squamous intraepithelial lesion development and oncogenesis by regulating KRAS expression in patients with high risk-HPV infection", 2022, Cell Death Discovery, doi: 10.1038/s41420-022-01013-5.
6	Mouse lung	M. Li, L. Fu, B. Lv, Y. Xiang, H. Xiang, D. Xu, H. Zhao, "Arsenic induces ferroptosis and acute lung injury through mtROS -mediated mitochondria-associated endoplasmic reticulum membrane dysfunction", Ecotoxicol. Environ. Saf., 2022, doi: 10.1016/j.ecoenv.2022.113595.
7	Mouse lung	Z. Zeng, H. Huang, J. Zhang, Y. Liu, W. Zhong, W. Chen, Y. Lu, Y. Qiao, H. Zhao, X. Meng, F. Zou, S. Cai, H. Dong, "HDM induce airway epithelial cell ferroptosis and promote inflammation by activating ferritinophagy in asthma", FASEB J., 2022, doi: 10.1096/fj.202101977RR.
8	Soybean leaf	Y. Cheng, H. Zhang, W. Zhu, Q. Li, R. Meng, K. Yang, Z. Guo, Y. Zhai, R. Ji, H. Peng, D. Dou, M. Jing, “Ferroptosis induced by the biocontrol agent Pythium oligandrum enhances soybean resistance to Phytophthora sojae ”, Environ. Microbiol., doi: 10.1111/1462-2920.16248.
9	Mouse testis	Y. Zhao, H. Zhang, J. Cui, J. Wang, M. Chen, H. Wang, X. Li, J. Li, "Ferroptosis is critical for phthalates driving the blood-testis barrier dysfunction via targeting transferrin receptor", Redox Biol., 2022, doi: 10.1016/j.redox.2022.102584.
10	Mouse colon	F. Yang, Y. Chen, Y. Xiao, H. Jiang, Z. Jiang, M. Yang, M. Li, Y. Su, Z. Yan, Y. Lin, D. Li, “pH-sensitive Molybdenum (Mo)-based polyoxometalate nanoclusters have therapeutic efficacy in Inflammatory Bowel disease by counteracting ferroptosis”, Pharmacol. Res., 2023, doi: 10.1016/j.phrs.2023.106645.
11	Mouse lung sample	W. Tang, J. Qin, Y. Zhou, W. Wang, F. Teng, J. Liu, L. Yi, J. Cui, X. Zhu, S. Wang, J. Dong, Y. Wei, "Regulation of ferroptosis and ACSL4-15LO1 pathway contributed to the anti-asthma effect of acupuncture", Int. Immunopharmacol., 2023, doi: 10.1016/j.intimp.2022.109670.
12	Mouse heart	H. Hu, L. Li, H. Zhang, Y. Zhang, Q. Liu, M. Chen, J. Ning, Y. Pang, W. Hu, Y. Niu, R. Zhang, "Mechanism of YY1 mediating autophagy dependent ferroptosis in PM2.5 induced cardiac fibrosis", Chemosphere, 2023, doi: 10.1016/j.chemosphere.2023.137749.
13	Mouse lung sample	B. Guo, Z. Zuo, X. Di, Y. Huang, G. Gong, B. Xu, L. Wang, X. Zhang, Z. Liang, Y. Hou, X. Liu, Z. Hu, "Salidroside attenuates HALI via IL-17A-mediated ferroptosis of alveolar epithelial cells by regulating Act1-TRAF6-p38 MAPK pathway", 2022, Cell Commun. Signaling, doi: 10.1186/s12964-022-00994-1.
14	Mouse testis	J. Ding, B. Lu, L. Liu, Z. Zhong, N. Wang, B. Li, W. Sheng, Q. He, “Guilu -Erxian-Glue alleviates Tripterygium wilfordii polyglycoside -induced oligoasthenospermia in rats by resisting ferroptosis via the Keap1/Nrf2/GPX4 signaling pathway”, Pharm. Biol., 2023, doi:10.1080/13880209.2023.2165114.
15	Mouse heart	J. Pan, W Xiong, A. Zhang, H. Zhang, H. Lin, L. Gao, J. Ke, S. Huang, J. ZHang, J. Gu, A. Chang, C. Wang, "The Imbalance of p53–Park7 Signaling Axis Induces Iron Homeostasis Dysfunction in Doxorubicin-Challenged Cardiomyocytes", Adv. Sci, 2023, doi:10.1002/advs.202206007.
16	3 x Tg-AD mouse brain	Xinlu Li, Jianfeng Chen, Wennuo Feng ,Chao Wang, Minyu Chen, Yifan Li, Jinghong Chen, Xinwei Liu, Qiong Liu, Jing Tian,“Berberine ameliorates iron levels and ferroptosis in the brain of 3 × Tg-AD mice”2023, PHYTOMEDICINE, doi:10.1016/j.phymed.2023.154962
17	Mouse testis	Lipeng Li , Zijie Pei , Ruiting Wu , Yaling Zhang , Yaxian Pang , Huaifang Hu , Wentao Hu , Zihan Geng , Tengfei Feng , Yujie Niu , Guimin Hao , Rong Zhang“FDX1 regulates leydig cell ferroptosis mediates PM2.5-induced testicular dysfunction of mice”，Ecotoxicology and Environmental Safety，2023，doi.org/10.1016/j.ecoenv.2023.115309

Q & A

Q Is it possible to use this kit with cell lysate?: A

No. The iron concentration in the cells is below the detection limit of the kit, meaning the cell samples cannot be measured.

Q How many samples can I measure?: A

Handling and storage condition

Handling and storage condition
Store at 0-4℃

SDS download

Iron Assay Kit -Colorimetric-

Iron Assay Kit -Colorimetric-