Rabbit anti p ulk1 ser555

HUVECs were lysed by RIPA buffer, and 30 µg of total protein was loaded and separated on 10% SDS-PAGE and transferred onto polyvinylidene difluoride (PVDF) membranes. After blocked with BSA, the PVDF membrane was incubated in primary antibody overnight at 4 °C (Rabbit anti-LC3B: 1:800, RRID: AB2137707; Rabbit anti-P62: 1:1000; Rabbit anti-cleaved-caspase 3: 1:1000, RRID: AB2070042; Rabbit anti-PARP: 1:1000, RRID: AB659884; Rabbit anti-ULK1: 1:1000, Rabbit anti-p-ULK1 ser-555: 1:1000, RRID: AB10707365; Cell Signaling Technology, Danvers, USA; Mouse anti-GAPDH:1:3000, Beyotime, Shanghai, China). GAPDH was used as a loading control. Signals were detected after incubation with horseradish peroxidase-conjugated secondary antibodies (1:5000, Bioworld, Shanghai, China) for 1 h at room temperature. The images were scanned by Fusion solo 4 (Vilber, Paris, France), and analyzed using Quantity One software 4.2.3 (Bio-Rad, USA).