Western blot was performed using Anti-SP1 Recombinant Rabbit Monoclonal Antibody (JF0950) (Product # MA5-32497) and a 95 kDa band corresponding to SP1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2), HEK-293 (Lane 3), HT-29 (Lane 4), PANC-1 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).