CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	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.

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

Autophagy detection was performed using CYTO-ID® Autophagy Detection Kit (Enzo Life Sciences) following the manufacturer’s protocol. In brief, cells were: control untreated or treated with DMSO, 500 nM rapamycin, or 500 nM rapamycin plus 50 µM chloroquine for 18 hours. The next day cells were stained with green detection reagent. Unstained cells were used as negative control. Samples were analyzed in green FL1 channel of flow cytometer (Beckman Coulter Inc). Arithmetic means were then used for analysis. Assay was repeated at least three times.

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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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