Western blot was performed using PKA C-alpha Polyclonal Antibody (Product # PA5-28160) and a 41 kDa band corresponding to PKA C-alpha along with a non-specific band at ~ 60 kDa was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), Jurkat (Lane 3), Hep G2 (Lane 4), and tissue extracts (30 µg lysate) of Mouse Brain (Lane 5), Rat Brain (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. 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, 1:10,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).