Overview

  • Product nameAnti-AIF antibodySee all AIF primary antibodies ...
  • Description
    Rabbit polyclonal to AIF
  • Tested applicationsWB, IHC-P, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Peptide corresponding to amino acids 517 to531 of human AIF. This sequence is identical tothose of mouse and rat AIF.

  • Positive control
    • K562 cell lysate
  • General notesApoptosis Inducing Factor

Properties

Applications

Our Abpromise guarantee covers the use of ab1998 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB Use a concentration of 0.25 - 1 µg/ml. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).Can be blocked with AIF peptide (ab7870). Can be blocked with AIF (internal) peptide (human).
IHC-P Use at an assay dependent dilution.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionProbable 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 diseaseDefects 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 similaritiesBelongs 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 localizationMitochondrion 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
  • Database links
  • Alternative names
    • AIFM1 antibody
    • AIFM1_HUMAN antibody
    • Apoptosis inducing factor 1, mitochondrial antibody
    • Apoptosis inducing factor antibody
    • Apoptosis inducing factor, mitochondrion associated, 1 antibody
    • Apoptosis-inducing factor 1 antibody
    • CMTX4 antibody
    • COXPD6 antibody
    • Harlequin antibody
    • Hq antibody
    • mAIF antibody
    • MGC111425 antibody
    • MGC5706 antibody
    • mitochondrial antibody
    • PDCD 8 antibody
    • PDCD8 antibody
    • Programmed cell death 8 (apoptosis inducing factor) antibody
    • Programmed cell death 8 antibody
    • Programmed cell death 8 isoform 1 antibody
    • Programmed cell death 8 isoform 2 antibody
    • Programmed cell death 8 isoform 3 antibody
    • Programmed cell death protein 8 antibody
    • Programmed cell death protein 8 mitochondrial antibody
    • Programmed cell death protein 8 mitochondrial precursor antibody
    • Striatal apoptosis inducing factor antibody
    see all

Anti-AIF antibody images

  • All lanes : Anti-AIF antibody (ab1998) at 1 µg/ml

    Lane 1 : K562 cell lysate
    Lane 2 : Rat heart tissue lysate
    Lane 3 : Mouse heart tissue lysate


    Predicted band size : 67 kDa
  • Immunohistochemistry of AIF in human retina with anti-AIF (IN) at 10 µg/ml.
  • ab1998 staining AIF in human U2OS cells by Immunocytochemistry/ Immunofluorescence.
    Samples were fixed using 4% paraformaldehyde.

    Left image shows fixed cells labeled with ab1998 (red) and cyclophilin A (green).
    Right image shows U2OS cell 6 hours after induction of apoptosis by 200 nM staurosporine.
    Note translocated of AIF to the nucleus upon induction of apoptosis.
  • ICC/IF image of ab1998 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1998, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-AIF antibody (ab1998)

This product has been referenced in:
  • Ozaki T  et al. Inhibitory peptide of mitochondrial µ-calpain protects against photoreceptor degeneration in rhodopsin transgenic S334ter and P23H rats. PLoS One 8:e71650 (2013). IHC ; Mouse . Read more (PubMed: 23951212) »
  • Sasaki-Osugi K  et al. Nuclear ALG-2 protein interacts with Ca2+ homeostasis endoplasmic reticulum protein (CHERP) Ca2+-dependently and participates in regulation of alternative splicing of inositol trisphosphate receptor type 1 (IP3R1) pre-mRNA. J Biol Chem 288:33361-75 (2013). Read more (PubMed: 24078636) »

See all 11 Publications for this product

Product Wall

Thank you for contacting us.

I would suggested searching the potential of this as caspase-independent apoptosis marker in publications. There are many publications related to this protein. Please have a look in pubmed.

As the process ...

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ab1998 : http://www.abcam.com/ab1998, where the immunogen is described asa peptide corresponding to amino acids 517 to 531 of human AIF. This sequence is identical tothose of mouse and rat AIF.

ab32042 : http://www.abcam.com/ab32042...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Breast cancer tissue and adjacent normal)
Specification Breast cancer tissue and adjacent normal
Fixative Formaldehyde
Antigen retrieval step Other
Permeabilization No
Blocking step H2o2 of abcam secondary antibody Kit Expose Kit as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 37°C
Username

Mrs. Thushara Jayan Pillai

Verified customer

Submitted Oct 24 2011

Abreviews
Application Western blot
Sample Human Cell lysate - whole cell (SW1573 lung cancer cell line)
Loading amount 50 µg
Specification SW1573 lung cancer cell line
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 8% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 27 2011

Thank you for your email. In your prior message with the attachment containing the Western blot image, incubation for 1 hr at RT was mentioned but not overnight at 4C. I agree with you and I would expect that you would see some sort of signal with your...

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Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab1998. At this time I would like to make the following suggestions in order to help improve your results. I would suggest extending the incubation period with t...

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This antibody has not been tested for IHC, therefore we cannot guarantee results.

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