CYTO-ID® Autophagy detection kit

Protocol tips

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	After washing, resuspend cells in 0.5 ml of freshly diluted Cyto‐ID reagents and incubate at 37°C for 30 min

Protocol tips
After washing, resuspend cells in 0.5 ml of freshly diluted Cyto‐ID reagents and incubate at 37°C for 30 min

Publication protocol

The Cyto-ID fluorescent reagent specifically labels autophagic vacuoles and colocalizes with LC3 (Oeste et al., 2013). Thus, autophagosome formation in cells was
detected using the Cyto-ID Autophagy Detection Kit (ENZO Life Sciences, Inc., Farmingdale, NY, USA). Briefly, cells were trypsinized, pelleted by centrifugation, and washed twice in a phenol-red-free culture medium with 2% FBS. The cells were resuspended in 0.5 mL of freshly diluted Cyto-ID reagents and incubated at 37 °C for 30 min. The Cyto-ID fluorescence of cells was immediately analyzed using a Gallios flow cytometer and the Kaluza analysis software (Beckman Coulter, Inc., Brea, CA, USA). The percentage of cells with Cyto-ID staining was used to assess the formation of autophagosomes.

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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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