Description
Key facts
Detection method:Colorimetric,
Sample types:Serum, Urine, Other biological fluids, Heparin Plasma,
Assay type:Quantitative,
Sensitivity:> 8 µM,
Range:0.008 - 5 mM,
Assay time:1h 30m,
Assay Platform:Microplate reader
Product details:
Iron Assay Kit ab83366 provides a simple convenient means of measuring ferrous (Fe2+) and/or ferric (Fe3+) iron in biological samples.
How the assay works
This ferrozine assay kit / iron assay kit uses an improved variant of the ferrozine molecule (ferene) which has been shown to give more accurate results for biological iron levels; that are closer to the results seen with mass spectrometry.
In a ferrozine assay, free ferrous iron (Fe2+) reacts with the ferrozine molecule (in this case the ferene molecule) to produce a stable colored complex with absorbance that can be measured with a colorimetric plate reader. The ferene iron probe used in this assay kit has absorbance at 593 nm when complexed with ferrous iron (Fe2+).
In Iron Assay Kit ab83366, in addition to the ferene iron probe and iron standard, an acidic assay buffer, with pH less than 5.5, is provided to release iron from iron carrier proteins. A reducing agent is also provided to convert ferric iron (Fe3+) to form Fe2+.
Ferrous iron (Fe2+) can be measured alone, or by addition of the reducing agent the level of total iron (Fe2+ and Fe3+) can be measured. 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
Getting the best performance from ab83366
This assay is not suitable for use with samples containing EDTA or citrate.
Related and recommended products
Iron Assay Kit ab83366 is often used in combination with Lipid Peroxidation (MDA) Assay Kit
ab118970
and GSH/ GSSG Assay Kit
ab239709
in the study of ferroptosis.
To measure total iron binding capacity and iron levels in serum, we recommend Total Iron-Binding Capacity (TIBC) and Serum Iron Assay Kit
ab239715
For iron staining in tissues, we recommend Iron Stain Kit (Prussian Blue Stain)
ab150674
Other 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.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
Biological function summary
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.
Pathways
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.
