Anti-LC3 (Rat) pAb

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	Primary antibody dilution 1:1000

Protocol tips
Primary antibody dilution 1:1000

Publication protocol

Total and cytosolic extracts were obtained using a commercial kit (Active Motif, Carlsbad, CA, USA). The following antibodies were used for immunoblotting: rabbit polyclonal anti-LC3 antibody (1:1000) (Code n.PD014, MBL International Corporation, Woburn, MA, USA), mouse anti-p62/SQSTM1 antibody (1:250) (Code n.MAB8028, R&D Systems, Minneapolis, MN, USA),
and mouse anti-b-actin monoclonal antibody (1:5000) (Code n.A5441, Sigma-Aldrich) as an internal control. Anti-rabbit (1:2000) (Code n.7074, Chemicon, Temecula, CA, USA) and anti-mouse (1:10,000) (Code n.103001, BD Pharmingen, Franklin Lakes, NJ, USA) antibodies were used as peroxidase-conjugated secondary antibodies. Chemiluminescence was detected using luminol solution (ECL Plus, GE Healthcare Amersham, Milan, Italy). Immunoreactive bands were visualized with an exposure time of
15 minutes (Kodak XOMAT). LC3 conversion from LC3 formI (LC3-I, 18 kDa) to LC3 form II (LC3-II, 16 kDa), normalized by b-actin level, was our reported autophagy index.

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