• Product name
    Iron Assay Kit
  • Sample type
    Urine, Serum, Other biological fluids, Tissue Extracts, Cell culture media
  • Assay type
  • Sensitivity
    > 8 µM
  • Range
    8 µM - 400 µM
  • Assay time
    1h 00m
  • Product overview

    Abcam's Iron Assay Kit provides a simple convenient means of measuring Ferrous and/or Ferric ion in sample. In the assay, ferric carrier protein will dissociate ferric into solution in the presence of acid buffer. After reduction to the ferrous form (Fe2+), iron reacts with Ferene S to produce a stable colored complex and give absorbance at 593 nm. 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 20 nmol in 50 µl sample, or 8 µM to 400 µM iron concentration in various samples.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    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.

  • Tested applications
    Suitable for: Functional Studiesmore details



Our Abpromise guarantee covers the use of ab83366 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

This kit is not compatible with plasma samples.


  • 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 mouse muscle lysate showing quantity (micrograms) per microgram total protein

  • Iron measured in mouse liver lysate showing quantity (micrograms) per microgram total protein

  • 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.



This product has been referenced in:
  • Wang F  et al. Excessive Iron and a-Synuclein Oligomer in Brain are Relevant to Pure Apathy in Parkinson Disease. J Geriatr Psychiatry Neurol 29:187-94 (2016). Read more (PubMed: 26940028) »
  • Alvarez LA  et al. NADPH oxidase-derived H2O2 subverts pathogen signaling by oxidative phosphotyrosine conversion to PB-DOPA. Proc Natl Acad Sci U S A 113:10406-11 (2016). Functional Studies . Read more (PubMed: 27562167) »

See all 8 Publications for this product

Customer reviews and Q&As

I contacted the laboratories. They confirmed that the assay buffer provided wtih ab83366 Iron Assay Kit has a pH of 4.6. Sulphuric acid is not known to interfere with the assay.

However, we would suggest to ensure that the pH level...

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Though the assay is incompatible with chelating agents such as EDTA and citrate, it is compatible with heparin and should be effective for assays of plasma collected with heparin. We do not, however, have data that confirms this.

Unfortunately, this kit is not suitable for use with plasma samples due to the presence of iron binding transferrin in plasma leading to inaccurate measurements of free iron.

1. All components should be stored in aliquots according to individual use requirements at -20C to prevent freeze-thaw cycles.

2. The plate can be covered by a plate cover and aluminium foil to protect from light during incubation. The incubat...

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Since the specific probe is proprietary they didn't have a spectrum to share, but in their experience reading at 570 nm should work and may simply reduce the sensitivity slightly.

All spectrophotometers have a window within which they are eq...

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Detection of iron levels in hair samples

Average Good 4/5 (Ease of Use)
We used the iron assay kit to evaluate whether iron can be detected in hair shaft samples. In this case, we used a hair sample from a Rhesus primate, since we had a sufficient amount to test. The hair sample was milled into a fine powder, homogenized with assay buffer (4x volume) and centrifuged as directed. We initially tested various small amounts of hair (ranging from 1.8 to 12.1mg), given that these samples are often only available in limited quantities. The resulting absorbance was very close to the base nm of the 0 standard, and translated to -0.043 to 0.034nmoles per 50μl sample. We had used two sets of standard concentrations: the set of standards recommended in the protocol (0-10nmol/well) and a 1:10 dilution of those standards (0-1nmol/well). Both standard curves had an R-value better than 0.98, and the diluted standard curve was used since it best encompassed the resulting nm.
We repeated the assay with a larger amount of hair (55.9mg). In the increased sample we were able to detect 0.14-0.23nmole per 50μl sample, which was closer to the stated range of the kit (0.4-20 nmole/50μl). This indicates that it is possible to detect iron in hair shaft sample using this kit. However, given the relatively large amount of sample needed, it is not an ideal method for limited sample sizes. The kit itself was very easy to use.

Dr. Erilynn Heinrichsen

Verified customer

Submitted Nov 05 2013

When the iron content in cell culture medium is <0.1uM, Transferrin is typically added to allow cell growth. Usually iron content is in the range of hundreds of micromolar in mammalian cell culture medium. We have never encountered such low iron con...

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The buffer in this kit should be compatible with generic protein assays. We would prefer using the Bradford method. If the protein cannot be detected that way, we would recommend to have duplicates of the same number of cells. One of the samples can th...

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This is a common problem seen in liver and serum samples. Lipoproteins in the sample are the main culprits behind this turbidity. For this purpose, we would recommend that to add 5 µl/well of 1 M SDS (28.8% or 288 mg/ml of SDS) to all thei...

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Unfortunately, the assay buffer composition is considered as proprietary information. However, I can confirm that it is compatible with the mentioned method to measure proteins.

To order from New Zealand, please contact our authoris...

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