Anti-Alcohol Dehydrogenase/ADH1A Antibody Picoband®

The metabolic enzymes ADH1, ALDH2 and CYP2E1 are significantly up-regulated by alcohol, and further up-regulated by BGXJW administration. A – E The mRNA levels of ADH1, ALDH2, ALDH1, CYP2E1 in liver tissues were subjected to RT-qPCR analysis. E Levels of ADH1, ALDH2, CYP2E1 in liver lysates after indicated treatment were determined by western blot, F Liver tissue sections were subjected to immunohistochemistry analysis of CYP2E1. All data are expressed as the mean ± SEM (n = 3). nsP > 0.05, *P < 0.05 vs ctrl group; nsP > 0.05 vs EtOH group. Crtl contrl, EtOH ethanol model, BGXJW Bao-Gan-Xing-Jiu-Wan
Index in PubMed under a CC BY license. PMID: 37127639

Effects of WEATs on ADH1A/ADH1B/ADH1G expression in the liver of mice. Representative immunoblots (A) and IHC images (B) of ADH1A/ADH1B/ADH1G expression in the liver. Quantification of ADH1A/ADH1B/ADH1G expression by western blotting (C) and IHC (D) . Each value represents the mean ± SEM ( n = 10).
Index in PubMed under a CC BY license. PMID: 35677547

Western blot analysis of ADH1A using anti-ADH1A antibody (PB10029).
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 HCCT tissue lysates,
Lane 2: human HCCP tissue lysates,
Lane 3: rat liver tissue lysates.
Lane 4: rat kidney tissue lysates.
Lane 5: mouse liver tissue lysates.
Lane 6: mouse kidney tissue 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 rabbit anti-ADH1A antigen affinity purified polyclonal antibody (Catalog # PB10029) 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-rabbit 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ADH1A at approximately 40 kDa. The expected band size for ADH1A is at 40 kDa.

IHC analysis of ADH1A using anti-ADH1A antibody (PB10029).
ADH1A was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ADH1A Antibody (PB10029) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Flow Cytometry analysis of U20S cells using anti-ADH1A antibody (PB10029).
Overlay histogram showing U20S cells stained with PB10029 (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 rabbit anti-ADH1A Antibody (PB10029, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of ADH1A using anti-ADH1A antibody (PB10029).
ADH1A was detected in a paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ADH1A Antibody (PB10029) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of ADH1A using anti-ADH1A antibody (PB10029).
ADH1A was detected in a paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ADH1A Antibody (PB10029) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of ADH1A using anti-ADH1A antibody (PB10029).
ADH1A was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-ADH1A Antibody (PB10029) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.