Immunoblotting reagents

[³H] oleic acid (45.5 Ci/mM) was purchased from PerkinElmer Life Sciences (Boston, MA). All immunoblotting reagents were obtained from Bio-Rad, Corp (Hercules, CA). Enhanced chemiluminescence (ECL) reagents were procured from GE Healthcare Life Sciences (Pittsburgh, PA). Tablets of protease inhibitor mixture were obtained from Roche Applied Science (Indianapolis, IN). Albumin was purchased from Sigma Chemical Co. (St. Louis, MO). All other bio-chemicals were of analytical grade. Sprague–Dawley rats, 150–200 g, were obtained from Harlan (Indianapolis, IN). All procedures involving animals were performed according to guidelines provided in the Guide for the care and Use of Laboratory Animals (Eighth Edition, 2011, published by The National Academies Press, USA) and the guidelines of the University of Central Florida’s Institutional Animal Care and Use Committee (IACUC) and an IACUC approved protocol was strictly followed.

Carbon monoxide (CO) and nitric oxide (NO) gases were from Matheson-TriGas Inc. (Houston, TX). NO was prepurified by passing through a NaOH trap to remove nitrous and nitric acid contaminants. Sodium hydrosulfite (Na₂S₂O₄), imidazole, heme, δ-aminolevulinic acid, isopropyl-1-thio-β-d-galactopyranoside, ampicillin, kanamycin, chloramphenicol, and egg lysozyme were from Sigma (St. Louis, MO). Restriction enzymes and other DNA modifying enzymes were purchased from New England BioLabs (Beverly, MA). Oligonucleotides were obtained from Integrated DNA Technologies (Coralville, IA). Reagents for DNA extraction and purification were from Qiagen (Valencia, CA). Immunoblotting reagents were from Bio-Rad (Hercules, CA). Plasmid vectors pET43.1a and pET28b, and Escherichia coli strains Rosetta 2(DE3)pLysS and Rosetta-gami B(DE3), and anti-HisTag monoclonal antibody were from Novagen (Madison, WI). TALON metal affinity resin was purchased from BD Biosciences Clontech (Palo Alto, CA). Other chemicals were all of reagent grade.