LC3 Antibody (APG8B) (N-term)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Dilute primary Ab at 1:100 - Incubate primary antibody at 8°C overnight

Protocol tips
- Dilute primary Ab at 1:100 - Incubate primary antibody at 8°C overnight

Publication protocol

The cells were cultured in glass-bottom culture dishes with coverslips (MatTek, P35GCOL-0–10-C) to 50 to 80% confluence. After the culture medium was aspirated, the cells were rinsed with PBS twice, fixed with 3.7% paraformaldehyde at 25°C for 10 min, and permeabilized with 0.2% Triton X-100 in PBS at 25°C for 10 min. After washing with PBS, the cells were incubated with primary antibody at 8°C overnight. The cells were washed with PBS 3 times and incubated with secondary antibody conjugated with a fluorescent dye (Molecular Probes, T-6390 or T-6391) at 37°C for 1 to 2 h. After washing with PBS 3 times, fluorescent staining of the cells was visualized under a Zeiss LSM710 confocal microscope (Peabody, MA 01960) or a Nikon Eclipse TE2000 inverted fluorescent microscope (Melville, NY 11747).

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