Anti xod

Proteins from lysates were prepared and resolved by 12% SDS-PAGE at 100 V, transferred for 1.5 h at 250 mA onto Nitrocellulose membranes, and analysed by immunoblotting as described previously^{56 (link)}. The information for primary antibodies is as follows: anti-ferritin (cat# 69090), SdhA (cat# 137040), and SdhB (cat# 178423) from Abcam (Cambridge, MA, USA), anti-Xod (cat# 55156-1-AP), citrate synthase (cat# 16131-1-AP), aconitase 2 (Aco2) (cat# 11134-1-AP), Ndufs1 (cat# 12444-1-AP), Ndufs3 (cat# 15066-1-AP), Uqcrc1 (cat# 21705-1-AP), and Uqcrfs1 (cat# 1843-1-AP) from Proteintech Group Inc. (Chicago, IN, USA), anti-Actin (cat# BM0627) from Boster (Wuhan, China), anti-Tubulin (cat# T0198) from Sigma-Aldrich (St. Louis, MO, USA), anti-TfR1 antibody (cat# 136800) from Zymed (San Francisco, CA, USA), anti-IscU, Fech, Irp1, and Irp2, anti-Fxn (self-made)^{31 (link)}, anti-CytC (cat# 1896-1) from Epitmics (Burlingame, CA, USA), anti-Vdac (cat# 4661S) from Cell Signaling Technology Inc (Shanghai, China).

Xanthine oxidase (XOD, EC 1.1.3.22) from cow milk (1 unit/mg protein), potassium oxonate (PO) and hypoXanthine (HX) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, United States). Xanthine was obtained from Dalian Meilun Biotech Co., Ltd. (Dalian, China). Allopurinol was obtained from Hebei Bailing Weichao Fine Materials Co., Ltd. (Langfang, China). Febuxostat (Feb) was bought from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China). BBR and DHB (Figure 1) were obtained from Chengdu HerbPurity CO., Ltd. (Chengdu, China). Assay kits for determination of SUA, blood urea nitrogen (BUN), creatinine, XOD and adenosine deaminase (ADA) were obtained from Nanjing Jiancheng Biotechnology Institute (Nanjing, China). Enzyme-linked immunoabsorbent commercial cytokine assay kits for murine TNF-α, IL-1β, IL-6 and IL-18 were purchased from Shanghai Chengshao Biotech Co., Ltd. (Shanghai, China). Mouse primers for ASC, caspase-1, NLRP3, IL-1β, URAT1, GLUT9 and XOD were purchased from Shanghai Sangon Biological Engineering Co., Ltd (Shanghai, China). Antibodies against NLRP3, ASC, IL-1β, and caspase-1 were obtained from Affinity Biosciences (Cincinnati, OH, United States). Anti-URAT1. anti-GLUT9 and anti-GAPDH were purchased from Proteintech Group, Inc. (Wuhan, China), and anti-XOD was purchased from Abcam (Cambridge, MA, United States).