Iron Assay Kit (Colorimetric)

• Biochemical assay

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Biochemical assay - Iron Assay Kit (Colorimetric) (AB83366)

Diagram showing the principles of the Iron assay method.

• Biochemical assay

PubMed

Biochemical assay - Iron Assay Kit (Colorimetric) (AB83366)

Wang et al. used Iron Assay Kit ab83366 and Lipid Peroxidation Assay Kit ab118970 to investigate ferroptosis in a mouse model of intestinal ischemia-reperfusion.

Ferroptosis occurs in a mouse model of intestinal I/R. Total iron, Fe2+, Fe3+, MDA, total glutathione, GSH, GSSG, and GSH/GSSG levels were evaluated in reperfusion tissues (n = 8).

To simulate intestinal ischemia-reperfusion (I/R), mice were anesthetized with 5% isoflurane and disinfected with Aner iodine. A midline incision was made in the upper abdomen to expose and dissociate the mesenteric artery. Closure of the mesenteric artery was performed for 60 min with a small artery clip to simulate the ischemic process. The arterial clamp was released to simulate the reperfusion process. Three different reperfusion times, namely, 15 min, 30 min, and 60 min, were used.

The total iron, Fe2+ and Fe3+ levels were assessed with an iron test kit (ab83366, Abcam) with a microplate reader at OD 593 nm. Malondialdehyde (MDA) was assessed with a Lipid Peroxidation Assay kit (ab118970, Abcam) at OD 532 nm. Tissue samples and cell samples were prepared according to the manufacturer’s protocols.

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Functional Studies - Iron Assay Kit (Colorimetric) (AB83366)

Assay of soluble free iron from a soil sample (5 μL of 100 μL buffer into which 100 mg of soil had been stirred), 5 μL of FBS and 5 μL of a 100 μM sample of iron standard.

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PubMed

Functional Studies - Iron Assay Kit (Colorimetric) (AB83366)

Leal SM Jr et al examined if iron availability regulates fungal growth in an infection as fungal infection intiates an iron sequestration response. Mice given Fe-dextran (Fe-Dex) and deferoxamine (Defox) shows a higher fungal mass (Fungal dsRed) compared to vehicle treated mice over 48 hours. Iron content was quantified in mouse serum using Iron assay kit (ab83366).

Leal SM Jr et al., PLoS Pathog 9(7), fig 2c. doi: 10.1371/journal.ppat.1003436. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

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Functional Studies - Iron Assay Kit (Colorimetric) (AB83366)

Example of iron standard curve using ab83366.

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Lab

Functional Studies - Iron Assay Kit (Colorimetric) (AB83366)

Iron measured in human urine showing concentration (micromolar)

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Functional Studies - Iron Assay Kit (Colorimetric) (AB83366)

Assay of total iron(II + III) in off-the-clot human serum (50 μl of serum was added per well). Data are mean ± SEM from 2 independent replicates, performed in duplicate wells.

• Biochemical assay

PubMed

Biochemical assay - Iron Assay Kit (Colorimetric) (AB83366)

Deng et al. used Iron Assay Kit ab83366 to investigate iron levels in fatty liver.

Concentration of liver tissue Fe2+ and liver tissue Fe3+ between NC (normal diet) group and HFD (high fat diet) group (n = 5).

The iron concentration of liver tissue was detected by an Iron Assay Kit (No. ab83366, Abcam, UK), according to the instructions provided by the manufacturer. Briefly, a standard dilution was prepared in the standard wells (100 μl), and the sample wells (40 μl) were prepared with sample solution (the volume was adjusted to 100 μl/well using iron assay buffer). Then, 5 μl of iron reducer was added to each standard well. The concentration of Fe2+ in the liver tissue samples was determined by adding 5 μl of assay buffer to each sample. To determine the total iron (Fe2+ + Fe3+) concentration of the liver tissue samples, 5 μl of iron reducer was added to each sample. Then, the reagents were mixed and incubated for 30 min in an oven at 37 ℃. Next, 100 μl of iron probe was added into each well and the solution was mixed well and incubated for 60 min in an oven at 37 °C in the dark. Finally, the OD value was detected with a microplate meter (wavelength = 593 nm).

• Biochemical assay

PubMed

Biochemical assay - Iron Assay Kit (Colorimetric) (AB83366)

Cao et al. used Iron Assay kit ab83366, Lipid Peroxidation (MDA) Assay Kit ab118970 and GSH Assay kit ab239727 to investigate the effect and mechanism of exosomal miR-26a derived from bone marrow MSCs (BMSCs) on liver fibrosis.

Overexpression of BMSC-derived exosomal miR-26a accelerated ferroptosis in HSCs. Measurement of Fe2+, MDA, and GSH of LX2 cells treated with miR-26a mimic-Exo or NC mimic-Exo.

The level of Fe2+, malonaldehyde (MDA), and glutathione (GSH) was analyzed by using Iron Assay kit (Cat# : ab83366, Abcam, Cambridge, UK), Lipid Peroxidation (MDA) Assay kit (Cat# : ab118970, Abcam), and GSH Assay kit (Cat# : ab239727, Abcam) based on the operating manual.

• Biochemical assay

PubMed

Biochemical assay - Iron Assay Kit (Colorimetric) (AB83366)

Junhing et al. used Iron Assay Kit ab83366 with a model of myocardial ischemia/reperfusion injury in rats treated with diltiazem and (-) epicatechin.

The concentration of Fe2+ in rat cardiac tissues : Control; I/R; L-EPI (I/R + L-EPI : 1 mg/kg/day of EPI); H-EPI (I/R + H-EPI : 2 mg/kg/day of EPI), DIL (I/R + DIL).

The total iron levels were measured in heart tissue in each group using the Iron Assay kit (ab83366, Abcam, UK). Tissues homogenate was lysed in 4 volume of buffer and centrifuged at 16,000 Ч g for 10 min. 5 µL of iron reducer agent was added into 50 µL of samples for Fe2+ assay, and 100 µL of iron probe solution was added to the samples and incubated at 25° C for 60 min kept in dark place. The absorbance of samples at 593 nm was measured using a micro spectrophotometer (Nanodrop, Thermo Fisher Scientific, USA).

• Biochemical assay

PubMed

Biochemical assay - Iron Assay Kit (Colorimetric) (AB83366)

Tang et al. used Iron Assay Kit ab83366 and MDA assay kit ab233471 to investigate a ferroptosis resistance mechanism involving the upregulation of TXNDC12, a thioredoxin domain-containing protein located in the endoplasmic reticulum.

Iron assays indicated that erastin- or RSL3-induced iron accumulation in HL60 and K562 cells during ferroptosis was unaffected by the knockdown of TXNDC12. In contrast, the absence of TXNDC12 accelerated erastin- or RSL3-induced lipid peroxidation, as evidenced by the malondialdehyde (MDA) assay.

The concentration of MDA in cells or tissues was determined using an MDA assay kit (Abcam, ab233471) following the manufacturer's instructions. The kit provided MDA standard and all necessary buffers. The assay involved the following main steps : serial dilution of MDA standards and test samples; addition of 10 µl of MDA color reagent stock solution to each well of MDA standard; incubation at room temperature for 10-30 minutes; addition of 40 µl of reaction solution and further incubation at room temperature for 30-60 minutes; measurement of the end-point absorbance at OD 695-700 nm using an absorbance plate reader with path-check correction.

The relative level of free ferrous iron (Fe2+) in cells was determined using an iron assay kit (Abcam, ab83366) following the manufacturer's instructions. The kit provided the iron standard and all necessary buffers. The assay involved the following main steps : setting up reaction wells; adding 5 µL of iron reducer to each standard well; adding 5 µL of assay buffer to each sample; mixing and incubating the standards and samples at 37°C for 30 minutes; adding 100 µL of iron probe to each well containing the iron standard and test samples; mixing and incubating them at 37°C for 60 minutes, protected from light. The output was measured immediately using a colorimetric microplate reader at OD 593 nm.

• Schematic Diagram

Supplier Data

Schematic Diagram - Iron Assay Kit (Colorimetric) (AB83366)

Representative image of Iron Assay Kit (Colorimetric) ab83366

Components shown from left to right :

- Iron Reducer

- Iron Standard I

- Iron Assay Buffer

- Iron Probe

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.