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    products/elisa/human-frataxin-elisa-kit10-x-96-well-plate-ab201121.pdf

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Cell Biology Apoptosis Mitochondrial
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SimpleStep

Human Frataxin ELISA Kit (10 x 96 well plate) (ab201121)

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  • SDS
  • Protocol Booklet
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Other - Human Frataxin ELISA Kit (10 x 96 well plate) (ab201121)
  • 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

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 (10 x 96 well plate)
    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

    Abcam’s Frataxin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Frataxin protein in in human cell 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.

    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

    Please refer to protocols.
  • Components 10 x 96 tests
    10X Human Frataxin Capture Antibody 1 x 6ml
    10X Human Frataxin Detector Antibody 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 200ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 5ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 50ml
    Antibody Diluent 4BI 1 x 60ml
    Human Frataxin Recombinant Protein 1 x 2ml
    Plate Seals 10 units
    Sample Diluent NS (ab193972) 1 x 120ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 10 units
    Stop Solution 1 x 120ml
    TMB Development Solution 1 x 120ml
  • 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
    • 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

    Images

    • Other - Human Frataxin ELISA Kit (10 x 96 well plate) (ab201121)
      Other - Human Frataxin ELISA Kit (10 x 96 well plate) (ab201121)
      ELISA Protocol Summary
    • 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.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

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