Recombinant nfκb p65 protein

Quercetin (QUR) (Cat# Q4951) and Carboxymethyl Cellulose (CMC) (Cat# C5678-500G) were purchased from Sigma Aldrich (St. Louis, MO, USA). Colorimetric and ELISA kits to measure levels of Troponin-I (Cat# E4737), creatinine kinase-MB (CKMB) (Cat# E4608), interleukin-6 (Cat# K4145), superoxide dismutase (SOD) (Cat# E458), tumor necrosis factor-α (TNF-α) (Cat# K1052), reduced glutathione (GSH) (Cat. NO. K454), and malondialdehyde (MDA) (Cat# K454) were purchased from BioVision, CA, USA. ROS assay kit (Cat# E-BC-K138-F) was purchased from Elabscience, CA, USA. A recombinant NF-κB p65 protein (Cat# 31102) and ELISA kit to measure the nuclear activity of NF-κB p65 (Cat# 40096) were purchased from Active Motif, Tokyo, Japan. An ELISA kit to measure serum and tissue levels of ANG II (Cat# CSB-E04494r) was purchased from CUSABIO, TX, USA. ANG II cocktail inhibitor (Cat# 9000681) was purchased from Cayman Chemicals, USA.

Probenecid was from Enzo Life Sciences, l-tryptophan-d₃ and ¹³C₃-cysteine were from Toronto Research Chemicals (Toronto, Canada); interferon-γ was from BD Pharmingen (Franklin Lakes, NJ); recombinant NF-κB–p65 protein was from Active Motif (Carlsbad, CA); ¹³C₂,¹⁵N-GSH was from Sigma-Aldrich (St. Louis, MO); and Kyn-CKA [4-(2-aminophenyl)-4-oxobut-2-enoic acid] was custom-synthesized by Toronto Research Chemicals (ε_{390 nm} = 3871 M⁻¹ cm⁻¹). ¹⁵N,¹³C₂-GSH and ¹³C₃-cysteine conjugates of Kyn-CKA were generated by reacting 100 equivalents of thiol with Kyn-CKA in 5 mM ammonium bicarbonate (pH 8.0) for 1 hour at 37°C, followed by solid-phase extraction using HyperSep C18 cartridges (Thermo Fisher Scientific). L-Kyn and N-formylkynurenine stocks were treated with 35 mg of 3-mercaptopropyl–functionalized silica from SiliCycle (Quebec City, Canada) for 15 min at room temperature to remove potential Kyn-CKA traces, followed by 0.22-μm filtration. Organic solvents were LC-MS grade (Thermo Fisher Scientific), and all other chemicals were of analytical grade and obtained from Sigma-Aldrich unless specified.