Western blot was performed using Anti-PYCARD Polyclonal Antibody (Product # PA5-85810) and a 21kDa band corresponding to Apoptosis-associated speck-like protein containing a CARD was observed across the cell lines tested and tissues tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), THP-1 (Lane 2), HL-60 (Lane 3), K-562 (Lane 4), HEK-293 (Lane 5), HeLa (Lane 6), Jurkat (Lane 7), Mouse Spleen (Lane 8), Rat Spleen (Lane 9), Mouse Liver (Lane 10), Rat Liver (Lane 11) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).