AIF Antibody (YA636)

AIF Antibody (YA636) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to AIF.

AIF functions dually as an NADH oxidoreductase in mitochondrial metabolism and as an apoptosis regulator (by similar). Upon apoptotic stimuli, it translocates from the mitochondrial intermembrane space to the cytosol and nucleus, triggering caspase-independent cell death (by similar). The nuclear soluble form (AIFsol) induces chromosomal DNA fragmentation (parthanatos) and interacts with EIF3G to inhibit protein synthesis while activating caspase-7 to amplify apoptosis. AIF also regulates respiratory chain biogenesis by controlling CHCHD4 mitochondrial import. Its pro-apoptotic isoform exhibits NADH oxidoreductase activity without inducing nuclear apoptosis.

Free

O95831

Predicted band size: 67 kDa; Observed band size: 67/50 kDa

Protein A affinity purified.

Mitochondrion intermembrane space, Mitochondrion inner membrane, Cytoplasm, Nucleus, Membrane.

Non-conjugated

Unmodified

AB_3102798

Cell Biology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:5000; ICC: 1:50-1:200; IHC-P: 1:50-1:1000; FC: 1:50-1:100; IP: Use at an assay dependent concentration.

• 
  Western blot analysis of extracts from C6 (lane 2(20μg) , NIH/3T3 (lane 3(20μg) , C2C12 (lane 4(20μg),using AIF Antibody (HY-P80006). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of SK-OV-3 cells labeling AIF with AIF Antibody (HY-P80006) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with AIF Antibody (HY-P80006) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of Hela cells labeling Smad4 with Smad4 Antibody (HY-P80006) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with AIF Antibody (HY-P80006)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Rat liver tissue using AIF Antibody (YA636). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80006, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Rat liver tissue using AIF Antibody (YA636). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80006, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Flow cytometric analysis of 1X10^6 Hela cells labeling AIF Antibody(HY-P80006, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/1000 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

WB, ICC/IF, IHC-P, IP, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human AIF.AA range:502-551.