CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Stain the cells with CYTO-ID for 30 min at 37°C in the dark	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
Stain the cells with CYTO-ID for 30 min at 37°C in the dark
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

Autophagy activity was determined using the Cyto-ID Autophagy Detection kit (Enzo Life Sciences, Farmingdale, NY) as per manufacturer’s protocol. Briefly, cells were seeded in a 6 well plate, and the following day cells were starved using EBSS as previously described. After starvation cells were labeled with Cyto-ID and Hoechst 33342 dyes for 30 minutes at 37 °C and then washed in PBS. Images were capture with Eclipse Ti-U using the same exposure times for all images acquired (Nikon Instruments Inc. Melville, NY).

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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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