Enzyme linked immunosorbent assay elisa kits for tnf α and il 1β

Silybin and LPS (Escherichia coli: Serotype O55:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α and IL-1β were purchased from R&D Systems, Inc. (Minneapolis, MN, USA). Anti-actin (4970), anti-phosphorylated (p-)p65 (3033) and anti-p65 (8242) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-NLRP3 (ab4270) and anti-caspase-1 (ab179515) antibodies were purchased from Abcam (Burlingame, CA, USA). Anti-ASC (sc-514414) antibody was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-mouse CD3-FITC (11-0032), CD11b-PE (12-0112) and Gr1-APC (53-5931) antibodies were obtained from eBioscience (San Diego, CA, USA). Alexa Fluor 488 anti-mouse IgG (A-11001) was from Invitrogen, Thermo Fisher Scientific (Waltham, MA, USA). The DAB staining kit was from KeyGen (Nanjing, China). All other chemicals were obtained from Sigma-Aldrich.

Primers were synthesized by Shanghai Generay Biotech Co., Ltd (Shanghai, China). The restriction enzymes StuI and HindIII were purchased from New England Biolabs, Ltd. (Beijing, China). The Taq DNA polymerase was obtained from Takara Biotechnology (Dalian) Co., Ltd. (Dalian, China). The reverse transcription-polymerase chain reaction (RT-PCR) purification, gel extraction and plasmid miniprep kits were purchased from Axygen (Corning Inc., Corning, NY, USA). The expression vector, pSumo-Mut, was modified and obtained from Novobio Scientific (Shanghai, China). Escherichia coli and ArcticExpress™ competent cells (DH5α and DE3 cells, respectively) were also obtained from Novobio Scientific. The Ni²⁺-IDA-Sepharose CL-6B affinity column was from Novagen^®, Merck KGaA (Darmstadt, Germany). SUMO protease was purchased from Novobio Scientific, and the cell counting kit 8 (CCK8) was purchased from Dojindo Molecular Technologies, Inc. (Rockville, MD, USA). Lipopolysaccharide (LPS) was from Sigma-Aldrich (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α and IL-1β were obtained from R&D Systems China Co., Ltd. (Shanghai, China).