T aoc assay kit

Ginsenoside Rg1 (purity ≥98%) and Astragaloside IV (purity ≥98%) were purchased from ChengDu ConBon Bio-tech Co., Ltd. (Chengdu, China). The methane dicarboxylic aldehyde (MDA), catalase (CAT), glutathione peroxidase (GSH-PX) and total anti-oxidative capacity (T-AOC) assay kits for rats were bought from Jiancheng Bioengineering Institute (Nanjing, China). Rat blood urea nitrogen (BUN), serum creatinine (SCr), β₂-microglobulin (β₂-MG) and urinary creatinine (UCr) ELISA kits were also purchased from Jiancheng Bioengineering Institute. Streptozotocin (STZ) was bought from Sigma (St Louis, MO, USA). Polyclonal antibodies used for immunohistochemical analysis included TGF-β1 (Santa Cruz Biotechnology, Dallas, TX, USA), Smad2/3 (Cell Sig-naling, Danvers, MA, China), Smad7 (Bioss, Woburn, MA, USA) and CTGF (Bioss). Rat SP immunohistochemistry kits were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The primer sequences of TGF-β1, Smad7, CTGF and ACTB (listed in Table 1) used for real-time PCR analysis were bought from Sangon Biotech Co., Ltd (Shanghai, China). Real-Time PCR Reagents & Kits and Total RNA Extractor (Trizol) were bought from Thermo Fisher Scientific. The other reagents used were of analytical grade.

Thirty sprouts of each treatment and measure their length with micrometer. Membrane damage of mustard sprouts was assessed by electrolyte leakage as described by Dionisio-Sese and Tobita,^{16 (link)} and free amino acids were measured according to the method of Moore.^{17 (link)} The glutathione peroxidase (GSH-PX), and total antioxidant capacity (T-AOC) were determined using a GSH-PX Assay Kit (A005-1-2, Nanjing Jiancheng Bioengineering Institute, China), and a T-AOC Assay Kit (A015-1-2, Nanjing Jiancheng Bioengineering Institute, China) respectively, according to the manufacturer's instructions. The protein concentration of plant protein extraction was determined according to Bradford^{18 (link)} using bovine serum albumin as a standard.

Glutathione peroxidase (GP_X) activity, thioredoxin reductase (TR) activity, total superoxide dismutase (SOD) activity, total antioxidant capacity (T-AOC), malondialdehyde (MDA) content, and total protein content of the muscle samples were determined in accordance with the instructions of a GPx activity assay kit (colorimetric method), TR activity assay kit (colorimetric method), SOD activity assay kit (WST-1 cell proliferation reagent method), T-AOC assay kit (2,2′-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid method), MDA assay kit (thiobarbituric acid method), and total protein quantitative assay kit (Bradford method) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), respectively. The reactive oxygen species (ROS) content of the muscle samples was determined in accordance with the instructions of an ROS assay kit (enzyme-linked immunosorbent assay methods) (Shanghai Jianglai Biological Technology Co., Ltd., Shanghai, China).