Rabbit anti nlrp3

Lysates were obtained by lysing the cell monolayer with phenylmethanesulfonyl fluoride (PMSF) and radioimmunoprecipitation assay (RIPA) lysis buffer (1:100). Total protein was separated in 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred to polyvinylidene difluoride (PVDF) membranes. Proteins were detected after labeling with specific primary antibodies [rabbit anti-NLRP3 (1:500, Bioss, China), rabbit anti-ASC polyclonal antibody (1:500, Bioss, China), rabbit anti-Caspase-1 (1:1000, Abcam, USA), rabbit anti cleaved Caspase1 (Arg317)/P10 (1:1000, Affinity, USA), rabbit anti-IL-1β antibody (1:1000, Abcam, USA), rabbit anti-BAX antibody (1:10000, Proteintech, USA), and rabbit anti-Bcl2 antibody (1:1500, Proteintech, USA)] followed by HRP-coupled secondary antibodies. The signals were detected by the addition of the chemiluminescent substrate (ECL Western Blot Kit, CWBIO, China). The protein amount was determined using Image J software. The level of GAPDH (1:1500, Proteintech, USA) was used to normalize the signal intensities.