Mouse tnf α and il 6 elisa kits

Baicalein (purity 98.58%) was purchased from Chengdu MUST Bio-Technology Co. Ltd. (Chengdu, China). LPS from Escherichia coli 055: B5 and Dimethyl sulfoxide(DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) was got from Hyclone (Thermo Scientific, Waltham, MA, USA). Dulbecco's modified Eagle's medium (DMEM) was acquired from Gibco (Invitrogen, Carlsbad, CA, USA). RIPA buffer and phenylmethanesulfonyl fluoride (PMSF) were gained from Wuhan Boster Bio-Technology Co. Ltd. (Wuhan, China). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylthiazolium bromide (MTT) was purchased from Beyotime Bio-Technology Co. Ltd. (Shanghai, China). NO Griess reagent was purchased from Beyotime (Shanghai, China). Mouse TNF-α and IL-6 ELISA kits were obtained from MULTI SCIENCES (Hangzhou, China). Rabbit monoclonal antibody against iNOS and mouse monoclonal antibody against Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were acquired from Abcam (Abcam Limited, Cambridge, United Kingdom). Peroxidase-conjugated affinipure goat anti-rabbit IgG and peroxidase-conjugated affinipure goat anti- anti-mouse IgG were obtained from Zen BioScience Co. Ltd (Chengdu, China). All other reagents were analytical grade.

TNF‐α and interleukin (IL)‐6 levels in mouse serum were measured using mouse TNF‐α and IL‐6 ELISA kits (MultiSciences, China). IL‐6 levels in the cultured supernatants of HCAECs and THP1‐derived macrophages were detected using a Human IL‐6 ELISA Kit (R&D Systems). The melatonin metabolite 6‐sulfatoxymelatonin in the urine of mice was measured using a Melatonin‐Sulfate Urine ELISA kit (IBL, Germany). Melatonin metabolite 6‐sulfatoxymelatonin in the urine of KD patients was also measured using a Melatonin‐Sulfate Urine ELISA kit (IBL). All assays were performed according to the manufacturer's instructions.