Radioimmunoprecipitation assay ripa buffer

Tobramycin and kanamycin A were purchased from Tokyo Chemical Industry Co., Ltd. (Chuo-ku, Tokyo, Japan). Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, trypsin-ethylenediaminetetraacetic acid, PD98059, and BCA kit were procured from Thermo Fisher Scientific (Waltham, MA, USA). α-MSH, NaOH, MTT, and kanamycin B were obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against tyrosinase, TRP-1, TRP-2, and MITF were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against p-p38, p38, p-JNK, JNK, p-ERK, ERK, p-AKT, AKT, and β-actin were purchased from Cell Signaling Technology (Danvers, MA, USA). Radioimmunoprecipitation assay (RIPA) buffer, dimethyl sulfoxide (DMSO), enhanced chemiluminescence (ECL) kit, and 2× Laemmli sample buffer were obtained from Biosesang (Sungnam, Gyeonggi-do, Korea). SP600125 and SB203580 were purchased from Cayman Chemical (Ann Arbor, MI, USA) and Calbiochem (San Diego, CA, USA); 2-DOS was obtained from Toronto Research Chemicals Inc. (Toronto, Ontario, Canada). EZ-LDH cell cytotoxicity assay kit was purchased from DoGenBio Co.,Ltd (Guro, Seoul, Korea).

Roswell Park Memorial Institute (RPMI) medium, TNBS, lipopolysaccharide (LPS) purified from Escherichia coli O111:B were supplied from Sigma-Aldrich (USA). Radioimmunoprecipitation assay (RIPA) buffer used as lysis buffer was supplied from Biosesang (Korea). Enzyme-linked immunoassay (ELISA) kits were supplied from BioTechne (USA) for the detection of interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α), and IL-6. Antibodies for inducible NO synthase (iNOS), cyclooxygenase (COX)-2, p65 (NF-κB), p-p65 (phosphorylated-NF-κB), and β-actin were procured from Cell Signaling Technology (USA). The TMB-Blotting Solution was supplied from Thermo Fisher Scientific (USA).