Gapdh polyclonal antibody

Mouse islet β-TC-6 cells were purchased from TONGPAI (Shanghai) Biotechnology Co. Ltd. PCB118 was purchased from AccuStandard Inc. (New Haven, CT, USA). Cell Counting Kit-8 (CCK8), ROS detection kit, and DMSO were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Specific antibodies against NLRP3, caspase-1, and fetal bovine serum were purchased from Cell Signaling Technology (CST, Boston, MA, USA). Anti-caspase-1 antibody (EPR4321) and anti-IL-1 β antibody were purchased from Abcam (USA). GAPDH polyclonal antibody was purchased from BioWorld (USA). ELISA kits for IL-6, IL-18, and CCL-2 were purchased from Cheng Lin Biological (Beijing, China).

The total protein was extracted from the cultured chondrocytes according to the manufacturer’s protocols using an RIPA buffer (Beyotime, Shanghai, China). After denaturation, 30 μg of total protein was electrophoresed for immunoblot analysis. The blots were probed with primary antibodies directed against runx2 (Abcam, Cambridge, UK) overnight at 4 °C. GAPDH polyclonal antibody (Bioworld, Minneapolis, MN, USA) was used as a housekeeping reference. Peroxidase-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific, Waltham, MA, USA) or goat anti-mouse IgG (Bioss, Shanghai, China) antibodies were used to visualize proteins using Western blotting chemiluminescence luminol reagent on a GeneGnome XRQ Western Blotting Analysis System (Syngene, Frederick, MD, USA). Working concentrations for each antibody were determined empirically based on the recommended stock solutions. Image J was used to quantify the band intensities of proteins of interest in the experimental and control groups.