Overview

  • Product nameIron Assay Kit
  • Tests
    100 x 1 assay
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay typeQuantitative
  • 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 applicationsFunctional Studiesmore details

Properties

  • RelevanceIron 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.
  • Alternative names
    • Fe
    • Fe++
    • Fe+++

Applications

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 dilution.

Iron Assay Kit images

  • 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)

  • Example of a Sample time line using ab83366
  • Example of iron standard curve using ab83366

Protocols

References for Iron Assay Kit (ab83366)

This product has been referenced in:
  • Budak H  et al. Stimulation of gene expression and activity of antioxidant related enzyme in Sprague Dawley rat kidney induced by long-term iron toxicity. Comp Biochem Physiol C Toxicol Pharmacol 166:44-50 (2014). Read more (PubMed: 25038477) »
  • Leal SM  et al. Targeting iron acquisition blocks infection with the fungal pathogens Aspergillus fumigatus and Fusarium oxysporum. PLoS Pathog 9:e1003436 (2013). Read more (PubMed: 23853581) »

See all 5 Publications for this product

Product Wall

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)
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.
Username

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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Thank you for contacting us.


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 ...

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Thank you for your enquiry.

Question 1: Can one test ferrous and ferric ions separately?

Answer 1: This test can for ferrous and ferric ions separately.

Question 2: Can one use heat denatured muscle tissue for the sample? ...

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Thank you for your enquiry.

I can confirm you can use perfused liver samples with all these kits.

ab83366 Iron Assay Kit will detect chelatable iron.

The kits are suitable for all mammalian species.

I hope th...

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