Immunocytochemical analysis of eIF3b in MG-63 cells using a eIF3b monoclonal antibody (Product #MA5-36159). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1:1,000 dilution. The nuclear counter stain is DAPI.

Western blot analysis of eIF3b using a eIF3b monoclonal antibody (Product #MA5-36159). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody at a dilution of 1:500 was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: A431 cell lysate, Lane 2: 293T cell lysate