CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Grow the cells in EBSS medium (amino acid depleted) medium to induce autophagy. Immunofluorescence staining to visualize the autophagosome.	Incase of Low CYTO-ID dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time. Incase of High CYTO-ID staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment.

Protocol tips
Grow the cells in EBSS medium (amino acid depleted) medium to induce autophagy. Immunofluorescence staining to visualize the autophagosome.
Downstream tips
Incase of Low CYTO-ID dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time. Incase of High CYTO-ID staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment.

Publication protocol

OV202NTC and OV202Sh1 cells were seeded in the 4-well tissue culture chambered slides and treated with indicated concentrations of AACOCF3, PG545 or starved with EBSS in the presence or absence of bafilomycin. After treatment, media was removed and washed with 1xPBS and AVs were detected with a Cyto-ID autophagy detection kit (Enzo Life Sciences, cat# ENZ-51031). Images were captured using a Zeiss LSM 510 microscope.

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Manufacturer protocol

Download the product protocol from Enzo Life Sciences for CYTO-ID® Autophagy detection kit below.

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