Western blot was performed using Anti-IkB alpha Polyclonal Antibody (Product # PA5-17888) and a 36 kDa band corresponding to NF-kappa-B inhibitor alpha was observed across cell lines and tissue tested, along with down regulation of protein upon TNF alpha treatment in HeLa and LNCaP. Also, the protein expression was restored In LNCaP upon MG132 treatment. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with TNF alpha (20 ng/mL for 10 min) (Lane 2), NIH/3T3 (Lane 3), NIH/3T3 treated with TNF alpha (20 ng/mL for 10 min) (Lane 4), LNCaP (Lane 5), LNCaP treated with TNF alpha (20 ng/mL for 10 min) (Lane 6), LNCaP treated with TNF alpha (20 ng/mL for 10 min) followed by MG132 (7.5 µg/mL for 1 hr) (Lane 7), Mouse Thymus (Lane 8), Rat Liver (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).