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

Apoptosis induction and AVO development were quantified using an in situ Cell Death Detection Kit (Roche Applied Sciences, Basel, Switzerland) and Cyto-ID™ Autophagy Detection Kit (Enzo LifeSciences), respectively, and analyzed by flow cytometry. Cells were stained for 30 min at 37°C and then harvested, after which green (510–530 nm) fluorescence emission from 1 × 104 cells illuminated with blue (488nm) excitation light was measured using a fluorescent activated cell sorter. A total of 50,000 events were collected, and data analysis was performed using Kaluza flow cytometry software (Beckman Coulter, Fullerton, CA, USA).

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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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