Western blot was performed using anti-Phospho-Cardiac Troponin I (Ser23, Ser24) Polyclonal Antibody (Product # PA5-35411) and a 24 kDa band corresponding to Phospho-Cardiac Troponin I was observed in Mouse Heart but decreased upon lambda phosphatase treatment, also the expression was negative in Mouse Brain. Tissue extracts (30 µg lysate) of Mouse Heart (Lane 1), Mouse Heart treated with lambda phosphatase (5 µL/mL for 6 hours) and Mouse Brain (Lane 3) 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). Lane 1 was treated with 5 µL/mL of lambda phosphatase for 6 hours at room temperature. 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).