Overview

  • Product name
    Iron Assay Kit
  • Sample type
    Urine, Serum, Other biological fluids, Tissue Extracts, Cell culture media
  • Assay type
    Quantitative
  • 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.

Properties

Images

  • 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.

Protocols

References

This product has been referenced in:
  • Yu SY  et al. Clinical features and dysfunctions of iron metabolism in Parkinson disease patients with hyper echogenicity in substantia nigra: a cross-sectional study. BMC Neurol 18:9 (2018). Read more (PubMed: 29343241) »
  • Lee YS  et al. Hepatocyte toll-like receptor 4 mediates lipopolysaccharide-induced hepcidin expression. Exp Mol Med 49:e408 (2017). Read more (PubMed: 29217822) »

See all 13 Publications for this product

Customer reviews and Q&As

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.

Yes, you can definitely measure ferrous and ferric acid separately with this kit. Please follow the protocol for measuring the ferrous and the total iron in the samples. When you subtract the ferrous levels from the total level, you get the ferric leve...

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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...

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We do not typically recommend using RIPA buffer with our kits since it contains SDS which can denature proteins but for this assay it could work. We have not tested with RIPA buffer. We recommend using the assay buffer that comes with the kit.

I can confrim that the ab83366 Iron Assay Kit will measure the free ferrous and ferric forms of iron and not the iron complexed in Heme.

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 we do not have a spectrum to share, but in our experience reading at 570 nm should work and may simply reduce the sensitivity slightly.

The emission profile shows a sharp peak at 580-590 nm, can measur...

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Detection of iron levels in hair samples

Average Good 4/5 (Ease of Use)
Abreviews
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.
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Dr. Erilynn Heinrichsen

Verified customer

Submitted Nov 05 2013

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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Normal serum Iron ~10-40 µM. Cells such as HeLa or cervical cells typically contain 1-0.9 pg iron/cell so 2 x 106 cells lysed in ~ 250 ul of assay buffer (Fe MW 55.85) should have about 4 – 8 nmol Fe per 50 ul test sample-possibly less if iron tightly ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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