QuantiChrom™ Iron Assay Kit
Protocol SDS  Printed Protocols and SDSs are not provided with kits.

For quantitative determination of iron ions Fe3+ and/or Fe2+ and evaluation of drug effects on iron metabolism.

Key Features
Sensitive and accurate. Linear detection range 27 μg/dL (4.8 μM) to 1,000 μg/dL (179 μM) iron in 96-well plate assay.

Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 40 min. Can be readily automated as a high-throughput assay for thousands of samples per day.

Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Cuvette or 96-well plate assay.

Low interference in biological samples. No pretreatments are needed. Assays can be directly performed on serum samples.


Biological (e.g. serum) and environmental samples


30 min

250 tests

Detection Limit
27 μg/dL (4.8 μM)

Shelf Life
12 months

More Details
Iron level in blood is a reliable diagnostic indicator of various disease states. Increased levels of iron concentration in blood are associated with blood loss, increased destruction of red blood cells (e.g. hemorrhage) or decreased blood cell survival, acute hepatitis, certain sideroachrestic anemias, ingestion of iron-rich diets, defects in iron storage (e.g. pernicious anemia). Decreased levels of blood iron may result from insufficient iron ingestion from diets, chronic blood loss pathologies, or increased demand on iron storage as during normal pregnancy. Simple, direct and automation-ready procedures for measuring iron concentrations find wide applications in research, drug discovery and environmental monitoring. BioAssay Systems iron assay kit is designed to measure total iron directly in serum without any pretreatment. The improved method utilizes a chromogen that forms a blue colored complex specifically with Fe2+. Fe3+ in the sample is reduced to Fe2+, thus allowing the assay for total iron concentration. The intensity of the color, measured at 590nm, is directly proportional to the iron concentration in the sample.

1. I would like to know how serum samples should be prepared for use in the assay.

Sera are usually obtained by centrifugation of clotted blood (fresh blood standing for about 30 min at room temperature). Both, fresh serum or frozen serum can be assayed directly (50 µL sample + 200 µL working reagent).

2. Do DTT or IGEPAL/NP-40 in cell lysis buffer will have any influence on the assay?

Our data show that up to 4% NP-40 has no effect on the DIFE-250 assay. Up to 20 mM β-mercaptoethanol also had no effect on the DIFE-250. We did not test DTT, but we believe DTT would not interfere with this assay either.

3. Can you use DIFE-250 to measure iron in whole blood?

No, whole blood needs to be diluted to a degree that the normal serum iron concentration is below the detection limit of the assay. The assay is not suitable for measuring hemoglobin bound iron.

4. Please let us know the preparation for the liver tissue.

Tissue samples should be dried overnight at 106°C and weighed. Samples can then be solubilized in 6N nitric acid by heating at 100°C to release protein-associated iron. The solution is neutralized with NaOH, diluted in deionized water as necessary, and assayed for iron concentration using the DIFE-250 kits.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Zhang, Y et al (2010). Diphthamide biosynthesis requires an organic radical generated by an iron-sulphur enzyme. Nature 465(7300):891-6. Assay: Iron in Bacteria cell (Pubmed).

2. Raulfs, EC et al (2008). In vivo iron-sulfur cluster formation. PNAS 105(25):8591-6. Assay: Iron in Bacteria cell protein (Pubmed).

3. Bandyopadhyay, S et al (2008). A proposed role for the Azotobacter vinelandii NfuA protein as an intermediate iron-sulfur cluster carrier. J Biol Chem. 283(20):14092-9. Assay: Iron in Bacteria cell protein (Pubmed).

4. Shah, YM et al (2009). Intestinal hypoxia-inducible transcription factors are essential for iron absorption following iron deficiency. Cell Metab. 9(2):152-64. Assay: Iron in Mouse erythrocytes (Pubmed).

5. Sudarshan, S et al (2009). Fumarate hydratase deficiency in renal cancer induces glycolytic addiction and hypoxia-inducible transcription factor 1alpha stabilization by glucose-dependent generation of reactive oxygen species. Mol Cell Biol. 29(15):4080-90. Assay: Iron in Human tumor cell (Pubmed).

6. Anderson, ER et al (2011). Intestinal hypoxia-inducible factor-2alpha (HIF-2alpha) is critical for efficient erythropoiesis. J Biol Chem. 286(22):19533-40. Assay: Iron in Mouse serum (Pubmed).

7. Chen, H et al (2009). Changes in iron-regulatory proteins in the aged rodent neural retina. Neurobiol Aging. 30(11):1865-76. Assay: Iron in Rat Serum (Pubmed).

8. Zhu, W et al (2010). Genetic iron chelation protects against proteasome inhibition-induced dopamine neuron degeneration. Neurobiol Dis. 37(2):307-13. Assay: Iron in Human neuron cell (Pubmed).

9. Habel, ME, Jung, D (2006). c-Myc over-expression in Ramos Burkitt’s lymphoma cell line predisposes to iron homeostasis disruption in vitro. Biochem Biophys Res Commun. 341(4):1309-16. Assay: Iron in Human lymphoma cell (Pubmed).

10. Yokosho, K et al (2009). OsFRDL1 is a citrate transporter required for efficient translocation of iron in rice. Plant Physiol. 149(1):297-305. Assay: Iron in Plant rice (Pubmed).

11. Chen, H et al (2009). Dysfunction of the retinal pigment epithelium with age: increased iron decreases phagocytosis and lysosomal activity. Invest Ophthalmol Vis Sci. 50(4):1895-902. Assay: Iron in Rat serum (Pubmed).

12. Ringseis, R et al (2010). Low availability of carnitine precursors as a possible reason for the diminished plasma carnitine concentrations in pregnant women. BMC Pregnancy Childbirth.10:17. Assay: Iron in Human plasma (Pubmed).

13. Hamlin, F, Latunde-Dada, GO (2011). Iron bioavailibity from a tropical leafy vegetable in anaemic mice. Nutr Metab (Lond). 2011 Feb 3;8:9. Assay: Iron in Mouse serum (Pubmed).

14. Waheed, MM et al (2011). Some Biochemical Characteristics and Preservation of Epididymal Camel Spermatozoa (Camelus dromedarius). Theriogenology. 2011 Jul 13. Assay: Iron in Camel Epididymal fluid (Pubmed).

To find more recent publications, please click here.
If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

- Fast turnaround
- Quality data
- Low cost

Please inquire via email or call 1-510-782-9988 x 2.
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QuantiChrom™ Iron Assay Kit
Catalog No: DIFE-250
Price: $329    Qty:
For orders of 10 or more kits, please call 1-510-7829988x1 or email us for best pricing and/or bulk order.

Shipping: RT
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: 4°C

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