Enzyme linked immunosorbent assay elisa kits for il 6 and tnf α

Tissue culture reagents, such as Roswell Park Memorial Institute 1640 (RPMI1640) and fetal bovine serum, were purchased from Gibco BRL Co. (Grand Island, NY, USA). All chemicals were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Primary antibodies, including anti-iNOS, anti-COX-2, anti-pIκBα, anti-β-actin, anti-p65, and anti-PCNA, were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); all the primary antibodies were rabbit source. Anti-rabbit secondary antibodies were purchased from Millipore (Billerica, MA, USA). Enzyme-linked immunosorbent assay (ELISA) kits for IL-6 and TNF-α were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Hyclone (Logan, UT, USA). An EZ-Cytox cell viability assay kit was purchased from DAEIL Lab (Seoul, Korea). LPS, Tween 20, sodium nitrite, phenylmethylsulfonyl fluoride, NP40 cell lysis buffer, dimethyl sulfoxide, protease inhibitor cocktail, and Griess reagent were purchased from Sigma-Aldrich (St. Louis, MO, USA). An RNeasy kit was purchased from QIAGEN (Hilden, Germany). A PrimeScript 1st strand cDNA synthesis kit and SYBR premix were obtained from Takara Bio Inc. (Tokyo, Japan). Antibodies against iNOS and STAT1 were purchased from Cell Signaling Technology (Danvers, MA, USA), and an antibody against β-tubulin was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Opti-MEM I medium, Lipofectamine 2000, and goat anti-rabbit and anti-mouse IgG horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Invitrogen (Carlsbad, CA, USA). Enzyme-linked immunosorbent assay (ELISA) kits for IL-6 and TNF-α were purchased from R&D Systems (Minneapolis, MN, USA), and an ELISA kit for IFN-β was obtained from Pestka Biomedical Laboratories (Piscataway, NJ, USA).