Overview

  • Product name
    Mouse AIF ELISA Kit
    See all AIF kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Cell extract 8 5%
  • Sample type
    Cell culture extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    5.9 ng/ml
  • Range
    7.81 ng/ml - 500 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 86 % - %

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

    AIF in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of AIF protein in mouse cell extract samples.


    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.


    AIF is a dual-function protein working both to support normal oxidative phosphorylation and as a key apoptotic factor. Calpains ad cathepsins cleave AIF in the mitochondria and affect its release during death signaling. Once released, AIF forms complexes with pro-apoptotic proteins and is delivered to the nucleus where it takes apart in DNA cleavage and chromatin condensation. Alteration of AIF function leads to severe metabolic disorders.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

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

Associated products

Applications

Our Abpromise guarantee covers the use of ab235651 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

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

  • 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 305.9 ng/mL in Raw264.7 cell extract, 387.4 ng/mL in K562 cell extract, and 242.1 ng/mL in C6 cell extract.

Protocols

References

ab235651 has not yet been referenced specifically in any publications.

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