Western blot was performed using Anti-BID Recombinant Rabbit Monoclonal Antibody (JM11-14) (Product # MA5-32642) and a 20 kDa band corresponding to BID was observed across cell lines and decreased upon Staurosporine and also with Actinomycin D along with TNF Alpha treatment. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), A-431 treated with Staurosporine (1 µM for 4 hr) (Lane 2), A-431 treated with Staurosporine (1 µM for 8 hr) (Lane 3), A-431 treated with Actinomycin along with TNF Alpha (5 µg/mL and 10 ng/mL for 5 hr) (Lane 4), Hep G2 (Lane 5), Jurkat (Lane 6), MOLT-4 (Lane 7), HeLa (Lane 8), PC-3 (Lane 9) and SH-SY5Y (Lane 10) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).