Mouse anti inducible nitric oxide synthase

Dulbecco's modified Eagle's medium (DMEM), phosphate-buffered saline (PBS), Hank's balanced salt solution (HBSS), fetal bovine serum (FBS), normal goat serum (NGS), and antibiotic-antimycotic were purchased from Gibco (Grand Island, NY, USA). Trimethyltin chloride, 2,7-dichlorofluorescin diacetate (DCFH-DA), 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), Tween-20, Hoechst 33258, SB203580, SP600125, apocynin, and BAY11-7082 were from Sigma-Aldrich (St. Louis, MO, USA). Rabbit antibodies against p-JNK, p-p38, p-ERK, p-IκBα, p38, and IκBα were purchased from Cell Signaling Technologies, Inc. (Beverly, MA, USA). Rabbit anti-JNK, ERK, and NF-κB p65 antibodies were from Santa Cruz Biotechnology (Santa, CA, USA). Mouse anti-inducible nitric oxide synthase (iNOS) was from BD Biosciences (Franklin Lakes, NJ, USA). Mouse anti-actin was purchased from EMD Millipore (Billerica, MA, USA). Rat anti-CD11b was from AbD Serotec (Oxford, UK). The goat anti-mouse, -rabbit, and -rat IgG (HRP-conjugated) secondary antibodies were from Enzo Life Science (Farmingdale, NY, USA). All other chemicals were purchased from Sigma-Aldrich.

2′,7′-Dichlorodihydrofluorescein diacetate (DCF-DA) was obtained from Molecular Probe (Eugene, OR, USA). Lipopolysaccharide (LPS; E. coli 0111:B4), hydrogen peroxide (H₂O₂) and tert-butyl hydroperoxide (t-BOOH) were purchased from Sigma-Aldrich. Other reagents were purchased from Sigma-Aldrich unless stated otherwise. Cultured media, serum-free supplements and antibiotics were purchased from Gibco (Carlsbad, CA, USA). Tissue culture plastics were from BD Bioscience (San Jose, CA, USA). Primary antibodies and suppliers were: rabbit or mouse anti-βIII tubulin (Covance), mouse anti-ED1 (CD68, for activated and phagocytic microglia/macrophage, Serotec, England), rabbit anti-phospho PKC (CS9371,Cell signaling technology, CO, USA), mouse anti-inducible nitric oxide synthase (iNOS, BD Bioscience, CA, USA), and goat anti-actin and anti-HSP70 (Santa Cruz Biotech, Dallas, TX, USA).