Horseradish peroxidase hrp anti mouse

Protein concentration was determined using Bradford Protein Assay Kit (BioRad, Hercules, CA, United States). Fifteen micrograms of protein samples were run on a resolving 10% acrylamide/polyacrylamide gel and transferred to a PVDF membrane. All primary antibodies (anti-PSMA diluted 1:1,000; anti-PRMT2 diluted 1:500; anti-PARP diluted 1:1,000; anti-PARG diluted 1:1,000; anti-CPT1alpha diluted 1:1,000; anti-PGC1alpha diluted 1:1,000; anti-βactin diluted 1:5,000) from Cell Signaling Technology (Danvers, MA, United States) were incubated at 4°C overnight. After washing, membranes were incubated with the secondary antibodies (1: 5,000 diluted horseradish peroxidase (HRP)-anti-mouse (Sigma-Aldrich (St. Louis, MO, United States) and 1:1,000 diluted anti-rabbit IgG (Cell Signaling Technology (Danvers, MA, United States). The bands were visualized by a chemiluminescence reagent (Cell Signaling Technology). Gel imaging Chemidoc MP System (BioRad, Hercules, CA, United States) was used to visualize and examine the protein bands. Bands were quantified using Scion Image 4.0 (Scion Corporation, Chicago, Illinois, United States).