Anti mor

NAc tissue was homogenized in a lysis buffer [21 (link)] for determination of total protein concentration, according to Bicinchoninic Acid (BCA) Protein Assay Kit (Pierce, IL) using bovine serum albumin as standard. After, protein samples were separated by electrophoresis on a 10% polyacrylamide gel and electrotransferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, MA, USA). Non-specific binding sites were blocked, and membranes were rinsed in buffer and incubated with primary antibodies: anti-β-actin (1:50,000; Sigma-Aldrich, St. Louis, USA), anti-DAT (1:500; Santa Cruz Biotechnology), anti-D1R (1:500, Santa Cruz Biotechnology), anti-TH (1:500, Santa Cruz Biotechnology), anti-ΔfosB (1:1000, Santa Cruz Biotechnology), and anti-MOR (1:500, Santa Cruz Biotechnology), followed by anti-goat (1:1000; Santa Cruz Biotechnology, CA, USA) or anti-rabbit (1:20,000; Santa Cruz Biotechnology, CA, USA) IgG horseradish peroxidase conjugate. Immunocomplexes were visualized using Luminata (Millipore, USA) according to the manufacturer’s instructions. Film signals were digitally scanned (Chemidoc™ Imaging Systems) and then quantified using ImageJ software. β-actin was used as an internal control, so that data were standardized according to actin values.