Polyvinylidene fluoride pvdf transfer membrane

Madopar, which is a combination of levodopa and benserazide, was obtained from Shanghai Roche Pharmaceuticals Ltd. (Shanghai, China). Dopamine, DOPAC, HVA, 5-HT, NE, HIAA, and MPTP were purchased from Sigma-Aldrich (St. Louis, MO, USA). A bicinchoninic acid (BCA) protein determination kit was obtained from CoWin Biosciences, Beijing, China. Diaminobenzidine (DAB) staining, and Nissl staining kits were purchased from Beyotime (Nanjing, China). The Polyvinylidene fluoride (PVDF) transfer membrane was purchased from Millipore Corp. (Bedford, MA, USA). Primary antibodies were obtained from Affinity Biosciences, Cell Signal Transduction, Cincinnati, OH, USA. Table 2 shows the primary antibody specifications and dilutions.

Restriction endonucleases, T4 DNA ligase and Pfu DNA polymerase were purchased from Thermo Fisher Scientific. DNA oligonucleotides were from Microsynth AG (http://microsynth.ch). High-glucose Dulbecco’s Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from Thermo Fisher Scientific (Gibco) and Penicillin/Streptomycin from Lonza. Proteinase inhibitor cocktail, Calpain inhibitor I, G418 were obtained from Sigma-Aldrich. Bradford-reagent was from Bio-Rad. Polyvinylidene fluoride (PVDF) transfer membrane and chemiluminescent substrate (Immobilon ECL substrate) were from Millipore. PNGaseF was purchased from New England Biolabs. Phosphatidylinositol dependent phospholipase C (PI-PLC), and PrestoBlue reagent were obtained from Thermo Fisher Scientific. The following primary antibodies were used: anti-Sho rabbit polyclonal antibody (Abgent, AP4754b), anti-β-actin mouse monoclonal antibody (Sigma, A1978). Secondary antibodies used were: Horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit IgG were from Jackson ImmunoResearch (715-035-151) and Pierce Biotechnology (31460), respectively. All other reagents and chemicals were purchased from Sigma-Aldrich.