Rabbit anti cle caspase 1

Equal amount of protein was resolved on SDS-PAGE gels and transferred to PVDF membrane. After blocking, membranes were incubated with primary antibodies rabbit anti ASC (1:1000, cell signaling technology, USA), rabbit anti-Cle-Caspase-1 (1:1000, cell signaling technology, USA), rabbit anti-pro-Caspase-1 (1:1000, Abcam, Cambridge, MA, USA), rabbit anti-acid ceramidase antibody (1:1000, LSBio, USA) and rabbit anti-β-actin (1:10000, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C. After overnight incubation, membranes were washed and incubated with donkey anti-rabbit-HRP IgG (1:5000, Santa Cruz Biotechnology, Dallas, TX, USA) for 1 h at room temperature. Finally, bands were detected by chemiluminescence technique using LI-COR Odyssey Fc and the band intensity of target proteins were normalized to β-actin and calculated with Image J software version 1.44p (NIH, Bethesda, MD, USA).