Anti phospho akt clone d9e

Tumor tissue or P. gingivalis infected cells were lysed in RIPA lysis buffer containing protease and phosphatase inhibitors, and protein concentration was measured by Bradford (Bio-Rad, Hercules, CA, USA). Proteins were separated by SDS-PAGE and transferred to nitrocellulose membranes. Rabbit antibodies were from Cell Signaling (Cell Signaling, Danvers, MA, USA): anti-phospho-AKT (clone D9E), and anti-Akt (pan, clone C67E7). Primary rabbit antibodies were used at 2 µg/mL overnight, followed by secondary goat anti-rabbit IgG-HRP (Abcam, Cambridge, UK). ECL-chemiluminescence detection kit (Biological Industries) was used to detect the proteins, and images were captured using a Bio-Rad imaging system (Bio-Rad, Hercules, CA, USA). Total and phosphor-AKT levels were determined by densitometry. Detailed information about western blot can be found at Figures S5–S7 (Supplementary Materials).