Horseradish peroxidase hrp linked anti rabbit igg antibody

Dulbecco’s Modified Eagle’s medium (DMEM), WST-8 colorimetric reagent, and KCN were purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Fetal bovine serum (FBS) and Dialyzed FBS were purchased from Equitech-Bio Inc. (Kerrville, TX, USA) and Thermo Fisher Scientific Inc. (Waltham, MA, USA), respectively. Anti-Akt, Anti-phosphorylated Akt, anti-GRP78, and anti-β-actin antibodies were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA). Horseradish peroxidase (HRP)-linked anti-rabbit IgG antibody (GE Healthcare Life Sciences, Buckinghamshire, UK) was used as secondary antibody. Mito Check Complex Activity Assay Kit (Cayman Chemical, Ann Arbor, MI, USA) was used to evaluate the effect of compound 1 on the mitochondrial complex I–V. Oxygen consumption of cells was measured by Oxygen Consumption Rate (OCR) Assay Kit (Cayman Chemical, Ann Arbor, MI, USA). Rotenone, thenoyltrifluoroacetone (TTFA), carbonyl cyanide m-chlorophenylhydrazone (CCCP), antimycin A, and oligomycin A were obtained from Tokyo Chemical Industry Co., LTD. (Tokyo, Japan), Wako Pure Chemical Industries, Ltd. (Osaka, Japan), Sigma-Aldrich (St. Louis, MO, USA), LKT Laboratories, Inc. (St. Paul, MN, USA), and Cayman Chemical (Ann Arbor, MI, USA), respectively. Other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA) or Kishida Chemical Co., Ltd. (Osaka, Japan).