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    products/elisa/human-frataxin-elisa-kit-ab176112.pdf

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Cell Biology Apoptosis Mitochondrial
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SimpleStep

Human Frataxin ELISA Kit (ab176112)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (1)References (5)

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Other - Human Frataxin ELISA Kit (ab176112)
  • Raw data value for example of frataxin standard curve
  • Example of Frataxin standard curve
  • Titration of HeLa cell extract within the working range of the assay
  • Quantification of Frataxin expression in different cell lines
  • Quantification of Frataxin expression in different human patient samples
  • Sandwich ELISA - Frataxin Human SimpleStep ELISA™ Kit (ab176112)
  • Linearity of dilution for Recombinant Frataxin

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 2.1 pg/ml
  • Range: 9.88 pg/ml - 800 pg/ml
  • Sample type: Cell culture extracts, Tissue Extracts
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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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

    2.1 pg/ml
  • Range

    9.88 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

    Human Frataxin ELISA kit (ab176112) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Frataxin protein in human cell culture and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate human Frataxin with 1.1 pg/ml sensitivity.

    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

            -Single-wash protocol reduces assay time to 90 minutes or less
            -High sensitivity, specificity and reproducibility from superior antibodies
            -Fully validated in biological samples
            -96-wells plate breakable into 12 x 8 wells strips

    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.


    ASSAY SPECIFICITY This kit recognizes both native and recombinant human Frataxin protein in cell and tissue extract samples only. Serum, plasma, saliva, and urine samples have not been tested with this kit.

  • 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.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Frataxin Capture Antibody 1 x 600µl
    10X Human Frataxin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Human Frataxin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Other Chaperones
    • Signal Transduction
    • Metabolism
    • Vitamins / Minerals
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Signal transduction proteins ELISA kits
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Neuroscience
    • Diseases
  • Relevance

    Function: Promotes the biosynthesis of heme and assembly and repair of iron-sulfur clusters by delivering Fe2+ to proteins involved in these pathways. May play a role in the protection against iron-catalyzed oxidative stress through its ability to catalyze the oxidation of Fe2+ to Fe3+; the oligomeric form but not the monomeric form has in vitro ferroxidase activity. May be able to store large amounts of iron in the form of a ferrihydrite mineral by oligomerization; however, the physiological relevance is unsure as reports are conflicting and the function has only been shown using heterologous overexpression systems. Modulates the RNA-binding activity of ACO1.
  • Cellular localization

    Cytoplasmic and Mitochondrial
  • Alternative names

    • CyaY
    • FA
    • FARR
    • FRDA
    • Friedreich ataxia protein
    • FXN
    • X25
    see all
  • Database links

    • Entrez Gene: 2395 Human
    • Omim: 606829 Human
    • SwissProt: Q16595 Human

    Associated products

    • Alternative Versions

      • Human Frataxin Antibody Pair - BSA and Azide free (ab256593)

    Images

    • Other - Human Frataxin ELISA Kit (ab176112)
      Other - Human Frataxin ELISA Kit (ab176112)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

    • Raw data value for example of frataxin standard curve
      Raw data value for example of frataxin standard curve

      Raw data values are shown in the table

    • Example of Frataxin standard curve
      Example of Frataxin standard curve

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

    • Titration of HeLa cell extract within the working range of the assay
      Titration of HeLa cell extract within the working range of the assay

      Background subtracted data from duplicate measurements are plotted.

    • Quantification of Frataxin expression in different cell lines
      Quantification of Frataxin expression in different cell lines

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

    • Quantification of Frataxin expression in different human patient samples
      Quantification of Frataxin expression in different human patient samples

      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.

    • Sandwich ELISA - Frataxin Human SimpleStep ELISA™ Kit (ab176112)
      Sandwich ELISA - Frataxin Human SimpleStep ELISA™ Kit (ab176112)This image is courtesy of an anonymous Abreview

      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.

    • Linearity of dilution for Recombinant Frataxin
      Linearity of dilution for Recombinant Frataxin

      Recombinant Frataxin was measured in biological samples in a 2-fold dilution series

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (5)

    Publishing research using ab176112? Please let us know so that we can cite the reference in this datasheet.

    ab176112 has been referenced in 5 publications.

    • Belbellaa B  et al. High Levels of Frataxin Overexpression Lead to Mitochondrial and Cardiac Toxicity in Mouse Models. Mol Ther Methods Clin Dev 19:120-138 (2020). PubMed: 33209958
    • Britti E  et al. Frataxin-deficient neurons and mice models of Friedreich ataxia are improved by TAT-MTScs-FXN treatment. J Cell Mol Med 22:834-848 (2018). PubMed: 28980774
    • 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). PubMed: 29773347
    • Piguet F  et al. Rapid and Complete Reversal of Sensory Ataxia by Gene Therapy in a Novel Model of Friedreich Ataxia. Mol Ther 26:1940-1952 (2018). PubMed: 29853274
    • 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 . PubMed: 28009062

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Kit still works when accidentally set the shaker to 95 degrees as opposed to Room Temperature

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    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.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Melanie Pride

    Verified customer

    Submitted Mar 01 2016

    Human frataxin in mouse tissue

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    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.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 10 2014

    Question

    Why is sample diluent NS included?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 11 2015

    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

    Kevin Hanson

    Abcam Scientific Support

    Answered on Sep 11 2015

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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