CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Use dye for 20 minutes in dark at 37C	Incase of Low CYTO-ID Green 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 Green 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
Use dye for 20 minutes in dark at 37C
Downstream tips
Incase of Low CYTO-ID Green 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 Green 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

Autophagic vacuoles were analyzed using a Cyto-ID autophagy detection kit (Enzo Life Sciences) according to the manufacturer’s protocol. In brief, cells were stained with Cyto-ID autophagy detection dye and Hoechst 33342 (Life Technologies) for 20 min at 37°C. Images were taken under the microscope (Nikon). EM analysis of autophagy was performed as described previously (Zhao et al., 2012).

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Manufacturer protocol

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

Download manufacturer protocol

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