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    products/elisa/human-aif-elisa-kit-apoptosis-inducing-factor-1-ab184858.pdf

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Cell Biology Apoptosis Nucleus Chromatin Condensation
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SimpleStep

Human AIF ELISA Kit (Apoptosis-Inducing Factor 1) (ab184858)

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  • SDS
  • Protocol Booklet
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Other - Human AIF ELISA Kit (Apoptosis-Inducing Factor 1) (ab184858)
  • Example of human AIF standard curve in 1X Cell Extraction Buffer PTR.
  • Interpolated concentrations of native AIF in human HeLa cell extract based on a 66.6 µg/mL extract load and human heart tissue extract based on a 20 µg/mL extract load.

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 60 pg/ml
  • Range: 0.156 ng/ml - 10 ng/ml
  • Sample type: Cell culture extracts, Plasma, Serum, Tissue Extracts
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human AIF ELISA Kit (Apoptosis-Inducing Factor 1)
    See all AIF kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HeLa extract 5 1.9%
    Inter-assay
    Sample n Mean SD CV%
    HeLa extract 3 3.2%
  • Sample type

    Serum, Plasma, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    60 pg/ml
  • Range

    0.156 ng/ml - 10 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 103.2 102% - 105.6%
    Cell culture media 99 97.3% - 101%
    Hep Plasma 107 105.6% - 108.4%
    EDTA Plasma 105.3 104.1% - 107.3%
    Cit plasma 106.2 101.7% - 110%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human AIF (Apoptosis-Inducing Factor 1) ELISA kit (ab184858) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of AIF protein in human serum, plasma, cell and tissue extract samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate human AIF with 60 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.

  • Notes

    AIF functions both as NADH oxidoreductase and as regulator of apoptosis. In response to apoptotic stimuli, it is released from the mitochondrion intermembrane space into the cytosol and to the nucleus, where it functions as a proapoptotic factor in a caspase-independent pathway. In contrast, AIF functions as an antiapoptotic factor in normal mitochondria via its NADH oxidoreductase activity. The soluble form of AIF (AIFsol) is found in the nucleus induces 'parthanatos' i.e. caspase-independent fragmentation of chromosomal DNA. AIF interacts with EIF3G, and thereby inhibits the EIF3 machinery and protein synthesis, and activates casapse-7 to amplify apoptosis. AIF plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells. AIF binds to DNA in a sequence-independent manner.

    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 AIF 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 CPI - HAMA Blocker (ab193969) 1 x 6ml
    Human AIF Capture Antibody (Lyophilized) 2 vials
    Human AIF 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
    • Nucleus
    • Chromatin Condensation
    • Cell Biology
    • Apoptosis
    • Nucleus
    • Other
    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Apoptosis
    • Nuclear
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Chromatin condensation
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cell cycle proteins ELISA kits
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • Chromatin condensation
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Neuroscience
    • Processes
  • Function

    Probable oxidoreductase that has a dual role in controlling cellular life and death; during apoptosis, it is translocated from the mitochondria to the nucleus to function as a proapoptotic factor in a caspase-independent pathway, while in normal mitochondria, it functions as an antiapoptotic factor via its oxidoreductase activity. The soluble form (AIFsol) found in the nucleus induces 'parthanatos' i.e., caspase-independent fragmentation of chromosomal DNA. Interacts with EIF3G,and thereby inhibits the EIF3 machinery and protein synthesis, and activates casapse-7 to amplify apoptosis. Plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells. Binds to DNA in a sequence-independent manner.
  • Involvement in disease

    Defects in AIFM1 are the cause of combined oxidative phosphorylation deficiency type 6 (COXPD6) [MIM:300816]. It is a mitochondrial disease resulting in a neurodegenerative disorder characterized by psychomotor delay, hypotonia, areflexia, muscle weakness and wasting.
  • Sequence similarities

    Belongs to the FAD-dependent oxidoreductase family.
  • Post-translational
    modifications

    Under normal conditions, a 54-residue N-terminal segment is first proteolytically removed during or just after translocation into the mitochondrial intermembrane space (IMS) by the mitochondrial processing peptidase (MPP) to form the inner-membrane-anchored mature form (AIFmit). During apoptosis, it is further proteolytically processed at amino-acid position 101 leading to the generation of the mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis in a caspase-independent manner.
  • Cellular localization

    Mitochondrion intermembrane space. Mitochondrion inner membrane. Cytoplasm. Nucleus. Cytoplasm > perinuclear region. Proteolytic cleavage during or just after translocation into the mitochondrial intermembrane space (IMS) results in the formation of an inner-membrane-anchored mature form (AIFmit). During apoptosis, further proteolytic processing leads to a mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis. Colocalizes with EIF3G in the nucleus and perinuclear region.
  • Target information above from: UniProt accession O95831 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • AIFM1
    • AIFM1_HUMAN
    • Apoptosis inducing factor
    • Apoptosis inducing factor 1, mitochondrial
    • Apoptosis inducing factor, mitochondrion associated, 1
    • Apoptosis-inducing factor 1
    • CMTX4
    • COWCK
    • COXPD6
    • Harlequin
    • Hq
    • mAIF
    • MGC111425
    • MGC5706
    • mitochondrial
    • Neuropathy, axonal motor-sensory, with deafness and mental retardation
    • neuropathy, axonal, motor-sensory with deafness and mental retardation (Cowchock syndrome)
    • PDCD 8
    • PDCD8
    • Programmed cell death 8
    • Programmed cell death 8 (apoptosis inducing factor)
    • Programmed cell death 8 isoform 1
    • Programmed cell death 8 isoform 2
    • Programmed cell death 8 isoform 3
    • Programmed cell death protein 8
    • Programmed cell death protein 8 mitochondrial
    • Programmed cell death protein 8 mitochondrial precursor
    • Striatal apoptosis inducing factor
    see all
  • Database links

    • Entrez Gene: 51060 Human
    • Entrez Gene: 9131 Human
    • Omim: 300169 Human
    • SwissProt: O95831 Human
    • Unigene: 424932 Human
    • Unigene: 476033 Human

    Associated products

    • ELISA kits

      • Human AIF Profiling ELISA Kit (ab126583)

    Images

    • Other - Human AIF ELISA Kit (Apoptosis-Inducing Factor 1) (ab184858)
      Other - Human AIF ELISA Kit (Apoptosis-Inducing Factor 1) (ab184858)

      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.

       

    • Example of human AIF standard curve in 1X Cell Extraction Buffer PTR.
      Example of human AIF standard curve in 1X Cell Extraction Buffer PTR.

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

    • Interpolated concentrations of native AIF in human HeLa cell extract based on a 66.6 µg/mL extract load and human heart tissue extract based on a 20 µg/mL extract load.
      Interpolated concentrations of native AIF in human HeLa cell extract based on a 66.6 µg/mL extract load and human heart tissue extract based on a 20 µg/mL extract load.

      The concentrations of AIF were measured in duplicate and interpolated from the AIF standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AIF concentration was determined to be 4.106 ng/mL in HeLa cell extract and 9.364 ng/mL in human heart tissue extract.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

    Publishing research using ab184858? Please let us know so that we can cite the reference in this datasheet.

    ab184858 has not yet been referenced specifically in any publications.

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