CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Use Rapamycin treated cells as positive control.	Stain the cells with CYTO-ID for 30min 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.

Upstream tips
Use Rapamycin treated cells as positive control.
Protocol tips
Stain the cells with CYTO-ID for 30min 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

Cyto-ID is a proprietary reagent specifically labels autophagic vacuoles and co-localizes with light chain 3 (LC3). Cyto-ID Autophagy detection kit (Enzo Life Sciences, Farmingdale, NY, USA) was used according to the manufacturer’s protocol. The fluorescence was measured by a plate reader (BioTek, Winooski, VT, USA). Cyto-ID Green reagent staining showed the relative fluorescence intensity of cells was increased in a dose-dependent manner, indicating the occurrence of autophagy. Rapamycin was used as a positive control [39].

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - BT-474 using CYTO-ID® Autophagy detection kit from Enzo Life Sciences.

Manufacturer protocol

Download the product protocol from Enzo Life Sciences for CYTO-ID® Autophagy detection kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - BT-474 using CYTO-ID® Autophagy detection kit from Enzo Life Sciences. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.