Western blot analysis of GSDMD in different lysates (10 µg/Lane). Exposure time: 2 minutes; 10% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. Samples were incubated in GSDMD Monoclonal antibody (Product # MA5-44940) using a dilution of 1:500 in 5% NFDM/TBST at room temperature for 2 hours followed by Goat Anti-Mouse IgG - HRP secondary antibody at a dilution of 1:100,000 for 1 hour at room temperature. Lane 1: PC-3 cell lysate; Lane 2: NIH/3T3 cell lysate; Lane 3: THP-1 cell lysate. Predicted band size: 53 kDa. Observed band size: 53 kDa.