Iron Assay Kit ab83366 provides a simple convenient means of measuring ferrous (Fe2+) and/or ferric (Fe3+) iron in biological samples.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Colorimetric
Serum, Urine, Other biological fluids, Heparin Plasma
Quantitative
8 - 400 µM
1h 30m
> 8 µM
Iron Assay Kit ab83366 provides a simple convenient means of measuring ferrous (Fe2+) and/or ferric (Fe3+) iron in biological samples.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Colorimetric
Serum, Urine, Other biological fluids, Heparin Plasma
Quantitative
8 - 400 µM
1h 30m
Microplate reader
> 8 µM
Blue Ice
-20°C
-20°C
-20°C
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 37°C
- add Iron Probe and incubate for 60 min at 37°C
- analyze with microplate reader
Technical Note: This assay is not suitable for use with samples containing EDTA or citrate.
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.
Iron is a chemical element essential for various metabolic processes. It functions as a cofactor for proteins involved in oxygen transport and electron transfer. Iron exists in several forms including ferritin and hemosiderin and has a molecular mass of approximately 55.85 dalton. In human bodies iron is expressed mainly in red blood cells liver muscle tissues and is stored in ferritin complexes. For histological studies techniques such as iron staining and Perls Prussian blue are used to visualize iron in tissues. Additionally assays of iron and iron TIBC (Total Iron Binding Capacity) measure iron levels and binding affinity.
Iron plays a role in cellular respiration DNA synthesis and electron transport. It is essential in hemoglobin within red blood cells and myoglobin in muscles where it binds and transports oxygen. Iron is also part of the iron-sulfur cluster and heme complexes important for electron exchange in cells. The blue stain used in histology highlights areas rich in iron aiding in diagnosis and research. You can assess iron availability and transport through tests like iron + TIBC.
Iron is integral to oxygen transport and mitochondrial electron transport chain regulation. It is important in erythropoiesis and energy production. Iron works with proteins like transferrin and ferritin important for maintaining iron homeostasis. Cytochrome oxidase a heme-containing enzyme plays an essential role in electron transport linked with iron. Iron is also vital in modulating cellular oxidative stress pathways which help in managing reactive oxygen species.
Abnormal iron metabolism leads to conditions like anemia and hemochromatosis. In anemia deficiency of iron results in reduced oxygen transport due to low hemoglobin levels. Hemochromatosis on the other hand involves iron overload damaging organs like the liver and heart. The balance of iron is regulated by proteins including transferrin and ferritin which maintain necessary levels for cellular function and prevent toxic accumulation. The use of iron staining in histology assists in identifying iron-related pathologies.
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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).
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 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.
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