Anti asc antibody sc 22514

MPTP was obtained from Sigma-Aldrich or Toronto Research Chemicals. MPP⁺, LPS, ATP, nigericin, staurosporine, propodium iodide, MCC950, dopamine and glibenclamide were purchased from Sigma-Aldrich. Mito-TEMPO was obtained from ENZO Life Sciences. MitoSOX was purchased from Invitrogen. Anti-mouse caspase-1 (AG-20B-0042) and anti-NLRP3 (AG-20B-0014) antibodies were obtained from Adipogen. Anti-mouse IL-1β antibody was obtained from R&D Systems (AF-401-NA). Anti-ASC antibody (SC-22514) and anti-phospho-ERK antibody (SC-7383) were purchased from Santa Cruz. Anti-CD45-BV421 antibody was obtained from BD Biosciences. Anti-ERK antibody (4695) and anti-phospho-JNK antibody (9251) were purchased from Cell Signaling. Anti-JNK antibody (554285) was obtained from BD pharmingen.

Unless otherwise indicated, all materials for Western blot analysis were purchased from BIO-RAD (Hercules, CA, USA). Sup and Lys were separated by SDS-PAGE (10% or 16%) using running buffer and transferred onto a polyvinylidene difluoride membrane (PVDF; 10849 A, Pall Co., Port Washington, NY, USA) using transfer buffer. The membranes were blocked with 3% skim milk and probed overnight at 4 °C with anti-mouse IL-1β antibody (AF-401-NA, R&D Systems, Minneapolis, MN, USA), anti-caspase-1 antibody (AG-20B-0042, AdipoGen Co., San Diego, CA, USA), anti-NLRP3 antibody (AG-20B-0014-C100, AddipoGen Co.), anti-Asc antibody (sc-22514, Santa Cruz Biotechnology, Santa Cruz, CA, USA), or anti-actin antibody (sc-1615, Santa Cruz Biotechnology). Membranes were further probed with HRP-conjugated 2^nd anti-sera (sc-2020, sc-2005 or sc-2004, Santa Cruz Biotechnology) and visualized by Power-Opti ECL^TM solution (BioNote Co., Gyeonggi-do, Republic of Korea) and a Cooled CCD camera System (AE-9105 EZ-Capture II, ATTO Technology, Tokyo, Japan).