LC3B Antibody Kit for Autophagy

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Use the cells pretreated with 30 µM chloroquine disphosphate for 16 h as a positive control.	Incubate the fixed and permeabilized cells with primary antibody for 1hr followed by 1hr incubation with secondary antibody.

Upstream tips
Use the cells pretreated with 30 µM chloroquine disphosphate for 16 h as a positive control.
Protocol tips
Incubate the fixed and permeabilized cells with primary antibody for 1hr followed by 1hr incubation with secondary antibody.

Publication protocol

Autophagy was detected with an LC3B Antibody Kit for Autophagy (ThermoFisher Scientific) containing anti-LC3B antibodies and chloroquine diphosphate. Before staining with Mito Red, the uninfected cells were pretreated with 30 µM chloroquine disphosphate for 16 h and used as a positive control. After fixation and permeabilization, the cells were incubated with primary anti-LC3B Abs for 1 h. Next, secondary anti-rabbit antibodies conjugated with FITC were used (1 h in the dark).

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