Anti-ALDH2 Antibody (monoclonal, 6H2)

• Catalog Number: orb614090
• Category: Antibodies
• Description: Anti-ALDH2 Antibody (monoclonal, 6H2). Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG1
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human ALDH2, different from the related mouse sequence by two amino acids, and from the related rat sequence by one amino acid.
• UniProt ID: P05091
• MW: 56 kDa
• Tested applications: FC, ICC, IF, WB
• Application notes: Western blot, 0.1-0.5 μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500μg/ml
• Antibody Type: Primary Antibody
• Clone Number: 6H2
• 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.
• Alternative names: Aldehyde dehydrogenase, mitochondrial; ALDH class
• Note: For research use only

Flow Cytometry analysis of SiHa cells using anti-ALDH2 antibody. Overlay histogram showing SiHa 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-ALDH2 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 ALDH2 using anti-ALDH2 antibody. ALDH2 was detected in immunocytochemical section of U20S 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 2 µg/mL mouse anti-ALDH2 Antibody overnight at 4°C. DyLight488 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 ALDH2 using anti-ALDH2 antibody. ALDH2 was detected in immunocytochemical section of U20S 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 2 µg/mL mouse anti-ALDH2 Antibody overnight at 4°C. DyLight488 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.

Western blot analysis of ALDH2 using anti-ALDH2 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 30 ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates, Lane 2: human placenta tissue lysates, Lane 3: human HEK293 whole cell lysates, Lane 4: human HL-60 whole cell lysates, Lane 5: human SHG-44 whole cell lysates, Lane 6: human THP-1 whole cell lysates, Lane 7: human K562 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA 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-ALDH2 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 1:10000 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 ALDH2 at approximately 56 kDa. The expected band size for ALDH2 is at 56 kDa.

Western blot analysis of ALDH2 using anti-ALDH2 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 30 ug of sample under reducing conditions. Lane 1: rat liver tissue lysates, Lane 2: rat kidney tissue lysates, Lane 3: rat heart tissue lysates, Lane 4: rat PC-12 whole cell lysates, Lane 5: mouse liver tissue lysates, Lane 6: mouse Ana-1 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA 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-ALDH2 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 1:10000 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 ALDH2 at approximately 56 kDa. The expected band size for ALDH2 is at 56 kDa.