Enhanced chemiluminescence ecl plus western blotting reagent

alpha-Mangostin (≥98% purity) and memantine hydrochloride (≥98%) were purchased from Sigma Aldrich (St Louis, MO, USA). Hydrogen peroxide (H₂O₂, 30%), Annexin V & Dead Cell Assay Kit, and Muse™ Caspase-3/7 Kit were obtained from Merck Millipore (Darmstadt, Germany). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 2′,7′-dichlorodihydrofluorescin diacetate (H₂DCFDA) were purchased from Molecular Probes (Eugene, Oregon, USA). Monoclonal antibodies against BAX, BCL-2, SIRT1, SIRT3, FOXO3a, CAT, SOD2, and actin, as well as horseradish peroxidase (HRP)-conjugated goat anti-mouse and rabbit secondary antibodies, were purchased from Cell Signaling Technology (Beverly, MA, USA). Enhanced chemiluminescence (ECL) Plus Western blotting reagent was supplied from Amersham Biosciences (Piscataway, NJ, USA). All other chemicals and reagents were of analytical grade from Sigma Aldrich (St Louis, MO, USA).

Minimum essential medium (MEM), Ham’s F-12 medium, fetal bovine serum (FBS), penicillin and streptomycin were purchased from Gibco BRL (Gaithersburg, MD, USA). Mouse monoclonal anti-actin (catalog number 3700), rabbit polyclonal anti-calpain (catalog number 2539), anti-calpastatin (catalog number 4146) and horseradish peroxidase-conjugated goat anti-mouse IgG and anti-rabbit IgG antibody were supplied by cell signaling (Beverly, MA, USA). Enhanced chemiluminescence (ECL) plus western blotting reagent was purchased from Amersham Biosciences (Piscataway, NJ, USA). The human dopaminergic neuroblastoma (SH-SY5Y) cell line was obtained from American Type Culture Collection (Manassas, VA, USA). SH-SY5Y cells are a thrice cloned subline of bone marrow biopsy-derived line SK-N-SH. SH-SY5Y cell has dopamine-β-hydroxylase activity and express tyrosine hydroxylase. 8-Hydroxyquinoline (99%), clioquinol (≥95%), and nitroxoline (96%) were purchased from Sigma-Aldrich (St Louis, MO, USA).