Anti p akt rabbit polyclonal antibody

Western blot was conducted according to the previous method (Kim et al., 2010 (link)). Protein extracts from brain tissue were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Protein separation was performed using 10% polyacrylamide with 0.05% bis-acrylamide. Proteins were then transferred to nitrocellulose and the blots were probed with anti-PI3K mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-p-PI3K rabbit polyclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-Akt rabbit polyclonal antibody (1:1,000, Cell Signaling Technology), and anti-p-Akt rabbit polyclonal antibody (1:1,000, Cell Signaling Technology). Peroxidase anti-rabbit IgG (1:5,000, Vector Laboratories), and peroxidase anti-mouse IgG (1:1,000, Vector Laboratories) were used as the secondary antibodies. Immunoreactivity was detected by enhanced chemiluminescence (ECL) detection kit (Santa Cruz Biotechnology). Film auto-radiograms were exposed from 5 to 15 min.

Western blot analysis was performed according to the previous method (Cho et al., 2017 (link); Kim et al., 2010 (link)). Protein separation was performed using 10% polyacrylamide with 0.05% bis-acrylamide. Proteins were then transferred to nitrocellulose and the blots were probed with anti-PI3K mouse monoclonal antibody (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p-PI3K rabbit polyclonal antibody (1:1,000; Santa Cruz Biotechnology), anti-Akt rabbit polyclonal antibody (1:1,000; Cell Signaling Technology Inc., Beverly, Massachusetts, USA), anti-p-Akt rabbit polyclonal antibody (1:1,000; Cell Signaling Technology), anti-GSK-3β rabbit polyclonal antibody (1:1,000; Santa Cruz Biotechnology), and anti-p-GSK-3β rabbit polyclonal antibody (1:1,000; Santa Cruz Biotechnology). Peroxidase anti-rabbit IgG (1:5,000; Vector Laboratories) and peroxidase anti-mouse IgG (1:10,000; Vector Laboratories) were used as the secondary antibodies. Immunoreactivity was detected by enhanced chemiluminescence (ECL) detection kit (Santa Cruz Biotechnology).