Anti pparg

Protein quantities were analyzed by standard Western blotting technique. Briefly, 25-50 µg total proteins lysates were separated on Mini PROTEAN Precast Gels with 2 X Laemmli sample buffer (with 2.5% β-mercaptoethanol) to a final 30 µL volume. The proteins were transferred onto a PVDF-membrane and blocked with superblock solution and probed with anti-UCP1 (1:1000), anti-PGC1-a (1:1000), anti-aP2 (1:1000), and anti-Ppar-g (1:1000) from Cell Signaling Technology (Danvers, MA); anti-CHOP (1:500), anti-p62 (1:500) from Santa Cruz Biotechnology, and anti-Cebp-a (1:1000, Abcam) and anti-α Tubulin (1:5000, Abcam) for overnight at 4˚C. Anti-rabbit or anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibodies (Santa Cruz Biotechnology) was used at a dilution of 1:5000 for 1 hr. at room temperature. The binding of specific antibodies was visualized via exposure to a photographic film after treating with enhanced chemiluminescence system reagents (Fisher Scientific, USA). The film was scanned and the band densities were quantified by ImageJ (NIH) software. The results were expressed as a relative ratio of the target protein to reference protein.