Anti-AIF antibody [E20] (ab32516)

Overview

• Product nameAnti-AIF antibody [E20]See all AIF primary antibodies ...
• Description
  Rabbit monoclonal [E20] to AIF
• Tested applicationsWB, IHC-P, ICC, Flow Cyt, IP, IHC-Fr more details
• Species reactivity
  Reacts with: Mouse, Rat, Human
  
• Immunogen

  A synthetic peptide correspndonding to residues near the C terminus of Human AIF.

• Positive controlWB: K562 cell lysate. IHC-P: Human cervical carcinoma tissue.
• General notesProduced under U.S. Patent No. 5,675,063.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
• Concentration information loading...
• Clonality Monoclonal
• Clone numberE20
• IsotypeIgG
• Research Areas
  • Metabolism
  • Pathways and Processes
  • Redox metabolism
  • Oxidative stress

  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Apoptosis

  • Cancer
  • Invasion/microenvironment
  • Apoptosis
  • Chromatin condensation

  • Epigenetics and Nuclear Signaling
  • Cell cycle
  • Apoptosis
  • Nuclear

  • Cell Biology
  • Apoptosis
  • Nucleus
  • Chromatin Condensation

Applications

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
  modificationsUnder 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
  • Entrez Gene: 51060 Human
  • Entrez Gene: 9131 Human
  • Entrez Gene: 9131 Human
  • Entrez Gene: 9131 Human
  • Entrez Gene: 26926 Mouse
  • Entrez Gene: 83533 Rat
  • Omim: 300169 Human
  • SwissProt: O95831 Human

  • SwissProt: O95831 Human
  • SwissProt: O95831 Human
  • SwissProt: Q9Z0X1 Mouse
  • SwissProt: Q9JM53 Rat
  • Unigene: 424932 Human
  • Unigene: 476033 Human
  • Unigene: 240434 Mouse
  • Unigene: 203165 Rat

  see all

• Alternative names
  AIFM1 antibodyAIFM1_HUMAN antibodyApoptosis inducing factor 1, mitochondrial antibody

  Apoptosis inducing factor antibodyApoptosis inducing factor, mitochondrion associated, 1 antibodyApoptosis-inducing factor 1 antibodyCOXPD6 antibodyHarlequin antibodyHq antibodymAIF antibodyMGC111425 antibodyMGC5706 antibodymitochondrial antibodyPDCD 8 antibodyPDCD8 antibodyProgrammed cell death 8 (apoptosis inducing factor) antibodyProgrammed cell death 8 antibodyProgrammed cell death 8 isoform 1 antibodyProgrammed cell death 8 isoform 2 antibodyProgrammed cell death 8 isoform 3 antibodyProgrammed cell death protein 8 antibodyProgrammed cell death protein 8 mitochondrial antibodyProgrammed cell death protein 8 mitochondrial precursor antibodyStriatal apoptosis inducing factor antibody

  see all

Anti-AIF antibody [E20] images

• 

  Western blot - AIF antibody [E20] (ab32516)
  Anti-AIF antibody [E20] (ab32516) at 1/1000 dilution + K562 cell lysate
  
  Predicted band size : 67 kDa
  Observed band size : 67 kDa
  
• 

  Immunoprecipitation - Anti-AIF antibody [E20] (ab32516)
  AIF was immunoprecipitated using 0.5mg K562 whole cell extract, 5µg of Rabbit monoclonal to AIF and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
  The antibody was incubated under agitation with Protein G beads for 10min, K562 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
  Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32516.
  Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
  Band: 67kDa: AIF
• 

  Immunohistochemistry (Paraffin-embedded sections) - AIF antibody [E20] (ab32516)
  Ab32516, at a 1/500 dilution, staining AIF in paraffin embedded human cervical carcinoma tissue by Immunohistochemistry.
• 

  Immunocytochemistry/ Immunofluorescence - Anti-AIF antibody [E20] (ab32516)
  ICC/IF image of ab32516 stained HeLa cells. The cells were 4% PFA 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 (ab32516, 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.
  
  
• 

  Flow Cytometry-AIF antibody [E20](ab32516)
  Overlay histogram showing K562 cells stained with ab32516 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32516, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with methanol (5 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.