2.12. Western blot analysis

The fold change of MYCBP protein in CRC tissues and transfected cells was detected using western blotting. Protein lysates were extracted from CRC tissues and cell lines using lysis buffer (Beyotime), followed by quantification and separation on 10% SDS‐PAGE (Invitrogen). The proteins were then immunoblotted onto the PVDF membrane (Millipore) by using a primary antibody for human MYCBP (1:1000; Abcam) and GAPDH (1:10 000; Abcam). HRP goat anti‐rabbit IgG (1:20 000; Boster Biotechnology) was then added as a secondary antibody. Enhanced chemiluminescence (ECL) system was used for visualizing bands.