Lactate dehydrogenase ldh cytotoxicity assay kit

Ginsenosides Rc and C-Mc were obtained from Ambo Laboratory (Dacejeon, Korea). ELISA kits for MMPs (MMP-1 and MMP-3), IL-6, TGF-β1, procollagen type I, and VEGF were purchased from R&D Systems, Inc. (Minneapolis, MN, USA). DMEM, FBS, and penicillin/streptomycin were purchased from Gibco BRL (Aidenbac, Germany). Glutathione content was assayed using a GSH assay kit (Cayman Chemical Co, Ann Arbor, MI, USA), and the lactate dehydrogenase (LDH) cytotoxicity assay kit was purchased from Roche Diagnostics GmbH (Roche Diagnostics, Mannheim, Germany). Antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Cell Science (Canton, MA, USA). Solvents were purchased from Samchun Pure Chemicals (Korea) unless otherwise stated.

Lactate dehydrogenase (LDH) Cytotoxicity Assay Kit (Roche Applied Science) was used to measure LDH released from the cells. The assay was performed using the manufacturer’s instructions. The absorbance of each sample at a wavelength of 492/620 nm was measured using a microplate reader (Microplate Reader SH-9000Lab, Corona Electric) and used to quantify cell viability.

Icariin powder, ≥95% purity verified by HPLC, was obtained from Nanjing Ze Lang Medical Technology Co. (Nanjing, China). d,l-Homocysteine was purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Minimum essential medium (MEM), penicillin and streptomycin (P/S), fetal bovine serum (FBS), phosphate buffered saline pH = 7.4, (10×) (PBS), 0.25% trypsin-EDTA. Phosphatase inhibitors cocktail and poly-l-lysine hydrobromide were obtained from Sigma-Aldrich, Inc. Lactate dehydrogenase (LDH) cytotoxicity assay kit and protease inhibitor cocktail were from Roche Diagnostics (Mannheim, Germany). Rabbit polyclonal antibodies against p-Akt (Ser473), Akt, p-ERK, ERK, p-JNK (Thr183/Thr185), JNK, p-Tau (Ser400/Thr403/Ser404), Total Tau (Tau-5), mouse Tyr-tubulin and mouse anti-Ac-tubulin (Lys40) were purchased from Cell Signaling Technology (Beverly, MA, USA). Mouse anti-α-tubulin and phosphatase inhibitor cocktail were purchased from Sigma-Aldrich, Inc. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit and goat anti-mouse antibodies were from DAKO (Glostrup, Denmark). Biomax X-ray film, Amersham enhanced chemiluminescence (ECL) detection kit and Amersham ECL plus detection kit were from GE Healthcare (New York, NY, USA). RNeasy mini kit, RT² First Strand kit, RT² Profiler PCR Array and 2× RT² SYBR Green MasterMix was purchased from QIAGEN (Valencia, CA, USA).