Anti apc ab15270

Cells were lysed in RIPA buffer (Beyotime Institute of Biotechnology) and the protein concentration was measured by the bicinchoninic acid (Pierce™ BCA Protein Assay kit; Thermo Fisher Scientific, Inc.). Each protein sample (~40 µg) was separated on a SDS-PAGE 10% gel (Mini-Protean-3 type; Bio-Rad Laboratories) and then transferred to a PVDF membrane (Merck KGaA) for 30 min. The membrane was blocked with 5% non-fat dry milk in TBST solution for 1 h. TBST with 3% bovine serum albumin was used to dilute each of the antibodies, including rabbit anti-human anti-Bax (cat. no. ab53154), anti-APC (ab15270, Abcam), anti-cyclin D1 (cat. no. ab226977), anti-c-Myc (cat. no. ab39688) and β-actin (cat. no. ab8227) (all 1:1,000) polyclonal antibodies, and goat anti-rabbit IgG (1:2,000; cat. no. ab6721) (all Abcam). The membrane was incubated with primary antibody overnight at 4°C. Following incubation with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000; cat. no. ABIN101988; Antibodies Online) at room temperature for 1 h, the ECL system (Thermo Fisher Scientific, Inc.) was used to detect the signals. The expression levels were quantified by ImageJ 1.46r software, and β-actin was used as an internal control.