Gapdh 6c5 rabbit igg

Briefly, protein extraction was performed with RIPA buffer, diluted in Laemmli buffer, separated by electrophoresis using a Mini Protean system 3 Electrophoresis Cell (Bio-Rad, Hercules, CA, USA) [7] . The membrane was incubated with the primary antibody caspase-3 antibody (Cell Signaling Technology®), B-cell lymphoma 2 (BCL-2), peroxisome proliferator-activated receptors (PPARα) and Peroxisome Proliferator-Activated Receptor Gamma, Coactivator 1 (PGC-1α) antibodies (Santa Cruz Biotechnology, Inc.) overnight. After that, the membrane was washed and incubated with the secondary antibody anti-IgG-HRP (Cell Signaling Technology®). Finally, immunodetection was performed using chemiluminescence according to the manufacturer's instructions (SuperSignal West Pico Chemiluminescent Substrate, Thermo Scientific, USA). The nitrocellulose membranes were exposed to X-Omat AR X-ray film (Eastman Kodak Co., USA) for predetermined time periods for each protein studied. The antibody used was the normalization of GAPDH (6C5), rabbit IgG (Santa Cruz Biotechnology, Inc., Europe), at 1:5000 dilution. Quantitative analysis of the blots was performed by Scion Image software (Scion Corporation, Frederick, Maryland, USA, available at http://www.scioncorp.com/) as previously described [7, 24] .