Western blot was performed using Anti-HSP27 Recombinant Rabbit Monoclonal Antibody (155)(Product # MA5-29333) and a 25kDa band corresponding to HSP27 was observed across cell lines tested except HT-1080 which is reported to be negative. A few non specific bands was also observed. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), MDA-MB-231 (Lane 2), PC-3 (Lane 3), DU 145 (Lane 4), HeLa (Lane 5), LNCaP (Lane 6) and HT-1080 (Lane 7) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).