Iron Assay Kit (Colorimetric) (ab83366)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr 30 min
- Sample type: Other biological fluids, Serum, Urine
- Sensitivity: 8 µM
Overview
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Product name
Iron Assay Kit (Colorimetric)
See all Iron kits -
Detection method
Colorimetric -
Sample type
Urine, Serum, Other biological fluids -
Assay type
Quantitative -
Sensitivity
> 8 µM -
Range
8 µM - 400 µM -
Assay time
1h 30m -
Product overview
Iron Assay Kit ab83366 provides a simple convenient means of measuring ferrous (Fe2+) and/or ferric (Fe3+) iron in biological samples.
Ferric carrier protein will dissociate ferric into solution in the presence of the acid assay buffer. If the assay buffer has a neutral pH, iron will bind tightly to the iron carrier protein. However, under acidic conditions (pH less than 5.5), iron no longer has a binding affinity for the carrier protein and will be dissociated and released into the solution as iron/ferric ions, whose concentration you can measure using this kit. The Iron Assay Buffer in this kit has an acidic pH that enables this release of iron/ferric ions into the solution.
Free ferrous iron (Fe2+) reacts with Iron Probe to produce a stable colored complex with absorbance at 593 nm. Ferric iron (Fe3+) can be reduced to form Fe2+ enabling the measurement of total iron (Fe2+ and Fe3+). The level of ferric iron (Fe3+) is calculated by subtracting ferrous iron from total iron.
A specific chelate chemical is included in the buffer to block copper ion (Cu2+) interference.
The kit measures iron in the linear range of 0.4 to 10 nmol or 8 µM to 200 µM.
Iron assay protocol summary:
- add samples and standards to wells
- for Fe2+ assay add assay buffer only, for total iron (Fe2+ and Fe3+) add iron reducer
- incubate for 30 min at 37ºC
- add Iron Probe and incubate for 60 min at 37ºC
- analyze with microplate reader -
Notes
This product is manufactured by BioVision, an Abcam company and was previously called K390 Iron Colorimetric Assay Kit. K390-100 is the same size as the 100 test size of ab83366.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests 2000 tests Iron Assay Buffer 1 x 25ml 20 x 25ml Iron Probe 1 x 12ml 20 x 12ml Iron Reducer 1 x 700µl 20 x 700µl Iron Standard I 1 x 100µl 20 x 100µl -
Research areas
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Relevance
Iron is essential to nearly all known organisms. It is generally stored in the centre of metalloproteins, in the heme complex, and in oxygen carrier proteins. Inorganic iron also contributes to redox reactions in the iron-sulfur clusters of many enzymes, such as nitrogenase and hydrogenase. -
Alternative names
- Fe
- Fe++
- Fe+++
Images
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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).
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Iron measured in human urine showing concentration (micromolar)
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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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Example of iron standard curve using ab83366.
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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.
Datasheets and documents
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SDS download
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Datasheet download
References (234)
ab83366 has been referenced in 234 publications.
- Xiong L et al. Downregulation of TNFAIP1 alleviates OGD/R‑induced neuronal damage by suppressing Nrf2/GPX4‑mediated ferroptosis. Exp Ther Med 25:25 (2023). PubMed: 36561622
- Zhang K et al. Heat shock protein 27 deficiency promotes ferrous ion absorption and enhances acyl-Coenzyme A synthetase long-chain family member 4 stability to promote glioblastoma cell ferroptosis. Cancer Cell Int 23:5 (2023). PubMed: 36639654
- Pitts HA et al. SPINK2 Protein Expression Is an Independent Adverse Prognostic Marker in AML and Is Potentially Implicated in the Regulation of Ferroptosis and Immune Response. Int J Mol Sci 24:N/A (2023). PubMed: 37298647
- Daoud L et al. Elucidating the effect of iron acquisition systems in Klebsiella pneumoniae on susceptibility to the novel siderophore-cephalosporin cefiderocol. PLoS One 17:e0277946 (2022). PubMed: 36580460
- Ma X et al. Lidocaine attenuates hypoxia/reoxygenation-induced inflammation, apoptosis and ferroptosis in lung epithelial cells by regulating the p38 MAPK pathway. Mol Med Rep 25:N/A (2022). PubMed: 35244190