Anti-PPAR alpha/PPARA Antibody Picoband®

Western blot analysis of PPAR alpha/PPARA using anti-PPAR alpha/PPARA antibody (A00600-2).
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 Hela whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: rat liver tissue lysates,
Lane 5: mouse liver 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-PPAR alpha/PPARA antigen affinity purified polyclonal antibody (Catalog # A00600-2) 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 PPAR alpha/PPARA at approximately 52 kDa. The expected band size for PPAR alpha/PPARA is at 52 kDa.

Puerarin regulates hepatic fatty acid metabolism. (A–I) qRT-PCR detected the expressions of Cd36 , Fabp1 , Fabp4 , Acc1 , Fasn , Scd1 , Ppar-α , Cpt1α , and Pgc1α in each group of livers. (J–K) The expression of PPARα in liver was detected by Western blotting and q-PCR. (L–N) The expression of PPARα in AML-12 cells treated with puerarin (0, 20, and 40 μM) and PA (500 μM) was detected by Western blotting and q-PCR. * means p < 0.05, ** means p < 0.01, ***means p < 0.001. NFD, normal fat diet; HFD, high-fat diet; HFD200, high-fat diet with puerarin (200 mg/kg/day) intervention; HFD300, high-fat diet with puerarin (300 mg/kg/day) intervention.
Index in PubMed under a CC BY license. PMID: 39055493

FMO5/PPAR-α signaling pathway is involved in puerarin-regulated lipid metabolism. (A) Molecular interaction of puerarin in the catalytic site of FMO3/FMO5; (B) qRT-PCR detected the expressions of Fmo3/Fmo5 in each group of livers; (C–E) The expression of FMO5 in AML-12 cells which were treated by puerarin (0, 20, and 40 μM) and PA (500 μM) was detected by Western blotting and q-PCR. (F) qRT-PCR and Western blotting detected the expressions of FMO5 in liver. (G–H) FMO5 protein expression in liver was detected by Western blotting and quantification with ImageJ. * means p < 0.05, ** means p < 0.01, ***means p < 0.001. NFD, normal fat diet; HFD, high-fat diet; HFD200, high-fat diet with puerarin (200 mg/kg/day) intervention; HFD300, high-fat diet with puerarin (300 mg/kg/day) intervention.
Index in PubMed under a CC BY license. PMID: 39055493

Flow Cytometry analysis of HepG2 cells using anti-PPAR alpha/PPARA antibody (A00600-2).
Overlay histogram showing HepG2 cells stained with A00600-2 (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-PPAR alpha/PPARA Antibody (A00600-2, 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.