AIF/AIFM1 Antibody (monoclonal, 2I5)

SKU: orb547790

Description

Anti-AIF/AIFM1 Antibody (monoclonal, 2I5). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Images & Validation

• Tested Applications: FC, ICC, IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Application Notes:
  Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Immunofluorescence, 2μg/ml Immunocytochemistry/Immunofluorescence, 5μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells. Add 0.2ml of distilled water will yield a concentration of 500μg/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG1
• Clone No.: 2I5
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human AIF, identical to the related mouse and rat sequences.
• Molecular Weight: 70 kDa

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

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; Neuropathy, axonal motor-sensory, with deafness and mental retardation antibody; neuropathy, axonal, motor-sensory with deafness and mental retardation (Cowchock syndrome) 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

Flow Cytometry analysis of Raji cells using anti-AIF antibody. Overlay histogram showing Raji cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-AIF Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of AIF using anti-AIF antibody. AIF was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-AIF Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of AIF using anti-AIF antibody. AIF was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/mL mouse anti-AIF Antibody overnight at 4°C. DyLight®550 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of AIF using anti-AIF antibody. AIF was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-AIF Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of AIF using anti-AIF antibody. AIF was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-AIF Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of AIF using anti-AIF antibody. AIF was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-AIF Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of AIF using anti-AIF antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: A549 whole cell lysates, Lane 2: Raji whole cell lysates, Lane 3: PC-3 whole cell lysates, Lane 4: Hela whole cell lysates, Lane 5: Caco-2 whole cell lysates, Lane 6: HepG2 whole cell lysates, Lane 7: THP-1 whole cell lysates, Lane 8: PANC-1 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-AIF antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for AIF at approximately 70KD. The expected band size for AIF is at 70KD.

Western blot analysis of AIF using anti-AIF antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: rat kidney tissue lysates, Lane 2: rat liver tissue lysates, Lane 3: mouse spleen lysates, Lane 4: mouse testicular tissue lysates, Lane 5: mouse kidney tissue lysates, Lane 6: RAW264.7 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-AIF antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for AIF at approximately 70KD. The expected band size for AIF is at 70KD.

Quick Database Links

UniProt

O95831