Anti-LC3A/B antibody

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	Dilute primary Ab at 1:1000

Protocol tips
Dilute primary Ab at 1:1000

Publication protocol

Proteins were isolated with RIPA lysis and extraction buffers (Thermo Fisher Scientific, Hudson, USA), and the supernatant was collected after centrifugation. The concentration of total proteins in the supernatant was determined using the Bradford protein quantification method and then isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Primary antibodies against SQSTM1/P62 (Abcam, ab91526), LC3B (Abcam, ab128025), PDK1 (Abcam, ab13755), and mTOR1 (Abcam, ab25880) were purchased from Abcam and used in a 1:1000 dilution in 5% BSA. P-AKT antibody was purchased from Cell Signaling Technology and diluted at 1:1000 in 5% milk. Following incubation with a horseradish peroxidase-coupled secondary anti-mouse/rabbit (Tianjin Sungene Biotech, Tianjin, China), protein bands were visualized with ECL Blotting Detection Reagents (NCI4106, Thermo Pierce ECL). The intensities of protein bands were determined using ImageJ software.

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Manufacturer protocol

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