Superoxide dismutase sod assay kit

BA (CAS registry no. 21967-41-9; molecular formula, C₂₁H₁₈O₁₁; molecular weight, 446.36; batch number 110715-201318) was provided by the National Institute for Food and Drug Control, Beijing, China. Magnesium sulfate was supplied by Jingqiu Chemical Chemical Co., Ltd (Beijing, China). BA-Mg was given by Professor Liu Cuizhe, Institute of Traditional Chinese Medicine, Chengde Medical College. LPS (Escherichia coli serotype 055:B5) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Dexamethasone (DEX) was produced by Aladdin Biochemical Technology Co., Ltd (Shanghai, China). Myeloperoxidase (MPO), malonaldehyde (MDA), and superoxide dismutase (SOD) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Water used in all experiments was purified by a Milli-Q Biocel Ultrapure Water System (Millipore, Bedford, MA). Enzyme-linked immunosorbent assay (ELISA) kits for tumor necrosis factor- (TNF-) α, interleukin- (IL-) 1β, and IL-6 were obtained from R&D Systems, Inc. (Minneapolis, MN, USA). Primary antibodies against p-IκBα, IκBα, MyD88, NF-κB p65, p-NF-κB p65, and TLR4 were obtained from Cell Signaling Technology (Danvers, MA, USA). Dulbecco's modified Eagle's medium (DMEM) was purchased from Life Technologies (Carlsbad, CA, USA).

Fish oil was purchased from Swiss company (Melbourne, Australia), corn oil from Shandong Sanxing corn technology Co., Ltd. (Shandong, China), and Vit E (α-tocopherol) from Sigma-Aldrich (St. Louis, USA). Fish oil, corn oil and Vit E were all certified as endotoxin free. IL-1β, IL-6, C-reactive protein (CRP), and TNF-α ELISA kits were purchased from Boster Biological Technology Co., Ltd. (Wuhan, China). 8-epi-prostaglandin F2α (8-epi-PGF2α) ELISA kit was purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China). Lactate dehydrogenase (LDH) and superoxide dismutase (SOD) assay kits were procured from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

DMEM was purchased from Thermo Fisher Biochemicals Co., Ltd. (USA). Foetal bovine serum, PLA2 enzyme, and penicillin were purchased from Gibco (USA). An AMV one-step RT-PCR kit was obtained from TaKaRa (China). Kuhuang injection extract powder was provided by Changshu Leiyunshang Pharmaceutical Co., Ltd. (China). A peristaltic pump was purchased from Master-Flex (German), and a 0.22 μm filter (Millex-GV) (German) was used. Sodium Deoxycholate (SDC) was obtained from VETEC (USA), and disposable indwelling intravenous needles (BD type Y 22 G) (USA) were used. A cell counting kit-8 (CCK-8) kit was purchased from Dojindo (USA), and reactive oxygen species (ROS) detection kits were purchased from Shanghai Biyuntian (China). Malondialdehyde (MDA) assay kits, superoxide dismutase (SOD) assay kits, glutathione (GSH) assay kits, and TBA assay kits were purchased from Nanjing Jiancheng Institute of Bioengineering (China). Aspartate transaminase (AST) test kits, alanine transaminase (ALT) test kits, lactate dehydrogenase (LDH) test kits, and alkaline phosphatase (ALP) test kits were all obtained from Nanjing Jiancheng Institute of Biological Engineering (China). CPZ was purchased from MCE (MedChemExpress) (USA), and GW9662 was obtained from Selleck (USA).

Streptozotocin (STZ), cyanamide (CYA), wortmannin (Wor), atractyloside (Atr), and Alda-1 were purchased from Sigma (St. Louis, MO, USA). Ethanol (EtOH) was obtained from Bengbu New Chemical Reagent Factory, China. 10% fetal bovine serum was obtained from Zhe Jiang Tianhang Biological corporation, China. Lactate dehydrogenase (LDH), Malondialdehyde (MDA), and superoxide dismutase (SOD) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute, China. CCK-8 assay kit was from Shanghai Bestbio Life Technology, China. The primers used were as follows: for ALDH2 forward: 5′-GTG TTC GGA GAC GTC AAA GA-3′ and reverse 5′-GCA GAG CTT GGG ACA GGT AA-3′, the amplified fragment length was 187 bp; for Bcl-2 forward: 5′-CTG GTG GAC AAC ATC GCT CTG-3′ and reverse: 5′-GGT CTG CTG ACC TCA CTT GTG-3′, the amplified fragment length was 227 bp; for Bax forward: 5′-GGA TCG AGC AGA GAG GAT GG-3′ and reverse: 5′-GCT CAT TGC CGA TAG TGA TGA CT-3′, the amplified fragment length was 464 bp; for β-actin forward: 5′-GAT GGT GGG TAT GGG TCA GAA-3′ and reverse: 5′-GGC CAT CTC TTG CTC GAA GTC-3′, the amplified fragment length was 630 bp. Mouse anti-ALDH2, anti-Bcl-2, anti-Bax, and anti-β-actin monoclonal antibodies were purchased from Santa Cruz Biotechnology (CA). Goat anti-mouse secondary antibodies were from Boston Co., Ltd., Wuhan, China.

P. citrinopileatus fruiting bodies preserved at the Plant Protection Institute of Beijing Academy of Agricultural and Forestry Sciences (Beijing, China) were sun-dried and crushed to obtain fine powder. Standard monosaccharides (d-xylose, d-glucose, d-galacturonic acid, d-mannose, d-glucuronic acid, l-rhamnose, d-fructose, d-arabinose and d-galactose), oleic acid, DEAE-cellulose, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), as well as palmitate were purchased from Sigma-Aldrich (USA). Superdex-200 column was acquired from the General Electric Company (GE, USA), and HepG2 cells were bought from the American Type Tissue Culture Collection (Manassas, VA, USA). Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s minimum essential medium (DMEM), phosphate-buffered saline (PBS), penicillin, trypsin solution, and streptomycin were purchased from Invitrogen (USA). Protein, triglyceride (TG), alanine transaminase (ALT), aspartate transaminase (AST), and superoxide dismutase (SOD) assay kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu Province, China). The rest of the chemicals as well as solvents were of analytical reagent grades and were obtained from Peking Chemical Co. (Beijing, China).

Silybin, β-glucuronidase (type B-10 from bovine liver), sulfatase (type H-1 from Helix pomatia), Ko143, MK-571, quinidine, cyclosporin A, 5(6)-Carboxy-2′,7′-dichlorofluorescein diacetate (CDCFDA), Rhodamine 123 (Rho123), and naringenin (NG) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Baicalein and baicalin were purchased from MelonePharma (Dalian Melone Biology Technology Co., Ltd., Dalian, China). Polyethylene glycol 400 (PEG400) was purchased from TCI (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan). Cremophor^® EL was purchased from Aladdin (Aladdin Industrial Corporation, Shanghai, China). Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), and superoxide dismutase (SOD) assay kits were purchased from the Nanjing Jiancheng Bioengineering Institute (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β ELISA assay kits were purchased from eBioscience (Thermo Fisher Scientific Inc., Vienna, Austria). All other chemicals and reagents were of analytical grade or higher and were commercially available. Serial dilution of the stock solutions with DMSO was prepared to produce working solutions of all compounds. The portion of DMSO in the final reaction solution was less than 1%.