LC3B antibody

Protocol tips

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	- Dilute Primary Ab at-1:200 dilution

Protocol tips
- Dilute Primary Ab at-1:200 dilution

Publication protocol

Immunoblotting was performed as described.40 Briefly, for chloroquine and ZnCl2 experiment, L6 myoblasts in six-well plates were treated with starvation as described above or 50 μM chloroquine with or without 50 μM ZnCl2 or 50 μM ZnCl2 alone in complete medium. For the experiments using bafilomycin A1 with different concentration ZnCl2 experiments, L6 myoblasts in six-well plates were washed in serum-free medium for 2 h and the media was aspirated, 0 to 50 μM ZnCl2 with or without 100 nM bafilomycin A1 in fresh serum-free medium was added followed by 20 h incubation. Proteins from cell lysates were separated by SDS-PAGE, followed by western blotting with anti-LC3 antibody made in rabbit (1:1000) or anti-β-actin antibody made in mouse (1:2000). Secondary rabbit or mouse antibodies were horseradish peroxidase (HRP)-conjugated (1:5000), and the signal was detected by autoradiography using an enhanced chemiluminescence western blotting kit (Perkin Elmer, NEL 104001EA).

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