Anti nlrp3

The surgically removed brains of normal and scrapie infected rodents were xed with 4% paraformaldehyde at 4°C for 16 h, and then moved to an embedding box and rinsed under running tap water for 3 h. The xed brain tissues were serially dehydrated and soaked in xylene for 1.5 h. The soaked tissue was embedded, and the tissue slices were prepared according to the routine neuropathological protocol.
Western blots 10% of brain homogenates were separated by 12% SDS-PAGE and electronically transferred to nitrocellulose membranes with a Semi-dry facility. After blocking with 5 % nonfat-dried milk in TBS at 37°C for 1 h, the membranes were incubated at 4°C overnight with speci c monoclonal or polyclone antibody, including anti-NLRP3 (1:000 dilution, AG-20B-0014-C100, AdipoGen, Switzerland), anti-ASC (1:500 dilution, Santa Cruz, USA) and anti-Caspase1 (1:500 dilution, 06-503-I, Millipore, USA). After washing with TBST (containing 0.1 % Tween-20, pH7.6) for 4 times, the membranes were incubated with horseradish peroxidase-conjugated anti-mouse or anti-rabbit antibody (1:5000 dilution, 115-035-003/111-035-003, Jackson, USA). Immunoreactive signals were developed using an enhanced ECL kit (PE Applied Biosystems, Foster City, CA, USA). Images were captured by ChemiDocTM XRSC Imager (Bio-Rad, USA).