Western blot analysis of LC3B was performed by loading 20 µg of THP-1 whole cell lysates, from cells either left untreated (Lane 1) or treated with 100 nM Rapamycin (Lane 2) for 24 hours, per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with a DyLight 650-conjugated LC3B polyclonal antibody (Product # PA5-22937) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.