Ab 775808

The JAK2, STAT3, pJAK2, pSTAT3, and cleaved caspase-3 proteins of myocardial tissue were extracted as per kit indications. The protein concentration was quantified. Western blot analysis was performed as previously described (11 (link)). Primary antibodies against the following antigens were used: JAK2 (1:1,000, AB_2128522; Cell Signaling Technology, Danvers, MA, USA), STAT3 (1:1,000, AB_331269; Cell Signaling Technology), phospho-JAK2 (phosphor-Y1007 + Y1008, 1:1,000, AB_775808; Abcam, Cambridge, UK), phospho-STAT3 (Tyr705, 1:1,000, AB_1658549; Abcam), β-actin (1:3,000, Bioworld Technology, Minneapolis, MN, USA). The secondary antibodies were horseradish peroxidase-conjugated goat anti-rabbit/mouse IgG (1:10,000, Cell Signaling Technology). The membranes were detected with an Odyssey two-color infrared scanner (LI-Cor Biosciences, Lincoln, NE, USA).

For Western blot analysis, proteins from different groups were extracted and protein concentration quantified. Then, samples were used for routine Western blot analysis; the detailed protocol has been described previously.17 (link) Antibodies were replaced by the primary antibodies JAK2 (1:1,000, AB_2128522; Cell Signaling Technology, Danvers, MA, USA), phospho-JAK2 (phospho-Y1007 + Y1008, 1:1,000, AB_775808; Abcam, Cambridge, UK), STAT3 (1:1,000, AB_331269; Cell Signaling Technology), phospho-STAT3 (Tyr705, 1:1,000, AB_1658549; Abcam), cleaved caspase 3 (1:1,000, AB_2070042; Cell Signaling Technology), and β-actin (1:3,000; Bioworld Technology, Minneapolis, MN, USA). The secondary antibody was horseradish peroxidase conjugated to goat antirabbit/mouse IgG (1:10,000; Cell Signaling Technology). The membranes were developed using an Odyssey two-color infrared scanner (LI-Cor Biosciences, Lincoln, NE, USA).