Overview

  • Product name
    Human Frataxin ELISA Kit
    See all Frataxin kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    HeLa 9 2.3%
    Inter-assay
    Sample n Mean SD CV%
    HeLa 3 6.3%
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    1.1 pg/ml
  • Range
    1.1 pg/ml - 800 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 92 86% - 98%
    Cell culture media 97 93% - 101%
    EDTA Plasma 89 88% - 92%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Frataxin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Frataxin protein in human cell culture and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Frataxin is a 17 kDa nuclear-encoded mitochondrial protein. In humans the gene is localized on chromosome 9 and is highly conserved during evolution. The gene is expressed in every cell, although in varying levels in different tissues and during development. The specific function of frataxin is still unknown, but it has been shown to play a role in iron metabolism. Studies have demonstrated that the deletion of the frataxin gene in yeast results in iron accumulation in mitochondria and loss of respiration. Recombinant human frataxin has been shown to bind iron in vitro, and increased mitochondrial iron levels have been observed in patients with Friedreich's Ataxia (FRDA). FRDA is an autosomal recessive, progressive degenerative disease characterized by neurodegeneration and cardiomyopathy it is the most common inherited ataxia.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab176112 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background subtracted data from duplicate measurements are plotted.

  • Interpolated values of Frataxin are plotted for the indicated cell lines based on a extract load of 10 µg/mL.

  • Transformed B lymphocyte cells from Friedreich's Ataxia (FRDA) samples, due to a stable homozygous GAA repeat insertion, are compared to heterozygous carrier B lymphocyte cells and control B lymphocyte cells. B lymphocyte cell extracts were analyzed across a 7-point titration (0.1-100 µg/mL) and frataxin levels were interpolated from the standard curve. Average interpolated values of Frataxin are plotted.

  • Standard curve generated using Abcam's Frataxin Human SimpleStep ELISA™ Kit (ab176112) with human brain tissue homogenate mixed in different proportions with mouse brain tissue homogenate. ab176112 did not cross-react with mouse frataxin. Both kinetic and endpoint readouts were used sequentially for a larger dynamic range. ab110353, 12ng human frataxin full length protein was also used to extended the standard curve to 12ng/ml.

Protocols

References

This product has been referenced in:
  • Khdour OM  et al. Lipophilic methylene blue analogues enhance mitochondrial function and increase frataxin levels in a cellular model of Friedreich's ataxia. Bioorg Med Chem 26:3359-3369 (2018). Read more (PubMed: 29773347) »
  • Kemp KC  et al. Cytokine therapy-mediated neuroprotection in a Friedreich's ataxia mouse model. Ann Neurol N/A:N/A (2016). Sandwich ELISA ; Human . Read more (PubMed: 28009062) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Step 13.5 "Seal the plate and incubate for 1 hour at room temperature on a plate shaker set to 400rpm"

Typically, we use a tube heater/shaker that has a lid on top of it, and place the plate on top of the lid for shaking at 400rpm, with no temperature set.

This time we accidentally left the machine set to 95 degrees the entire 1 hour. The kit still worked accordingly.

Dr. Melanie Pride

Verified customer

Submitted Mar 01 2016

Question
Answer

Our entire SimpleStep ELISA™ range of products comes with a bulk packaged group of buffers, including the Sample Diluent NS. Thus, even though some of the kits do not require use of the Sample Diluent NS such as ab176112, we still send you the buffer because it is more efficient and cost-effective for us to do so and get the product to you quickly and at lower cost.

Read More

Human frataxin in mouse tissue

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Excellent performance and consistant results over multiples plates.
Does not cross-react with mouse frataxin
Used both kinetic and endpoint readout sequentially for a larger dynamic range.
I also purchased Ab110353 12 ng human frataxin full length protein and extended the standard curve to 12 ng/ml.
I recommend this product.
Attached is an example where human brain tissue homogenate was mixed in different proportions with mouse brain tissue homogenate.

Abcam user community

Verified customer

Submitted Nov 10 2014

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

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