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, Tissue Extracts, Urine
- Sensitivity: 8 µM
Product nameIron Assay Kit (Colorimetric)
See all Iron kits
Sample typeUrine, Serum, Other biological fluids, Tissue Extracts
Sensitivity> 8 µM
Range8 µM - 400 µM
Assay time1h 30m
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
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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests 2000 tests Iron Assay Buffer WM 1 x 25ml 20 x 25ml Iron Probe NM 1 x 12ml 20 x 12ml Iron Reducer Green 1 x 700µl 20 x 700µl Iron Standard (100 mM) Yellow 1 x 100µl 20 x 100µl
RelevanceIron 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.
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).
Iron measured in human urine showing concentration (micromolar)
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.
Example of iron standard curve using ab83366.
Assay of iron(II) and total iron(II+III) in perfused rat liver homogenate (an equivalent of 8 mg wet tissue was used per well). Data are mean ± SEM from 2 independent replicates, performed in duplicate wells.
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.
ab83366 has been referenced in 111 publications.
- Song Y et al. Human umbilical cord blood-derived MSCs exosome attenuate myocardial injury by inhibiting ferroptosis in acute myocardial infarction mice. Cell Biol Toxicol 37:51-64 (2021). PubMed: 32535745
- Wang K et al. Branched-chain amino acid aminotransferase 2 regulates ferroptotic cell death in cancer cells. Cell Death Differ 28:1222-1236 (2021). PubMed: 33097833
- Liu J et al. NUPR1 is a critical repressor of ferroptosis. Nat Commun 12:647 (2021). PubMed: 33510144
- Zhu W et al. A paradox: Fe2+-containing agents decreased ROS and apoptosis induced by CoNPs in vascular endothelial cells by inhibiting HIF-1a. Biosci Rep 41:N/A (2021). PubMed: 33345265
- Luo H & Zhang R Icariin enhances cell survival in lipopolysaccharide-induced synoviocytes by suppressing ferroptosis via the Xc-/GPX4 axis. Exp Ther Med 21:72 (2021). PubMed: 33365072