Rabbit anti phospho β catenin

Differentiation medium with Aβ40 peptide treatment for each condition was collected and analyzed by WB in order to determine the presence of the Aβ40 peptide in its monomeric form. For the detection of cell death, β-catenin and p-β-catenin, 50 µg of cellular extracts of differentiated cultures after Aβ peptide treatment were analyzed. Samples were boiled for 5 min, loaded on a 12% sodium dodecyl sulphate (SDS)-polyacrylamide gel, electrophoresed and transferred to nitrocellulose membranes (GE Healthcare). Membranes were blocked in PBS containing 5% nonfat dairy milk with 0.05% Tween20 (Sigma) for 1 h at RT. Blots were incubated overnight at 4 °C with primary antibodies against mouse β-actin (1:1000; Sigma), mouse anti-Aβ 4G8 (1:1000; Covance) mouse anti-phospho-Histone H2A.X (γH2AFX; 1:1000; Millipore (Burlington, MA, USA)), rabbit anti-β-catenin (1:1000; Cell Signaling) or rabbit anti-phospho-β-catenin (1:1000; Cell Signaling). The blots were developed using peroxidase-conjugated goat anti-rabbit (GARPO; 1:3000; Vector Laboratories), peroxidase-conjugated horse anti-mouse (HAMPO; 1:3000, Vector Laboratories) or goat anti-rabbit (GARPO; 1:3000; Vector Laboratories) for 1 h at RT and visualized using the ECL system (Millipore).