CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Supplement PBS with 0.2% B27. - Stain cells at 37°C for 30 min in the dark

Protocol tips
- Supplement PBS with 0.2% B27. - Stain cells at 37°C for 30 min in the dark

Publication protocol

To stain cells with Cyto-ID® (Enzo life science Icn.
NY, USA) we followed the manufacturer’s protocol with
modifications. In brief, cells were first washed with PBS
(supplemented with 0.2% B27) and then mixed with
either Cyto-ID® solution. The cells were then incubated
at 37°C for 30 min in the dark, then washed twice with
PBS to remove the free dyes. Cells were fixed in 4% para‐formaldehyde for 20 min at room temperature then cells were washed twice with PBS, followed by mounting with fluorescence mounting media (Dako, CA, USA). Images were acquired by the Carl Zeiss LSM 700 Meta confocal microscope and analyzed using Carl Zeiss ZEN image software. Stained samples were analyzed by flow cytometric analysis using FACS CaliburTM (Becton Dickinson Biosciences, CA, USA). All experiments were performed three times in duplicate.

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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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