CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Add reagent and incubated cells for 30 minutes 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
- Add reagent and incubated cells for 30 minutes 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

AML-12 and RAW 264.7 cells were seeded at a density of 4×105 cells/dish. The cells were treated with fresh medium alone, control vehicle alone (0.05% DMSO, v/v) or Sch B (0.1, 1, 10, and 25 μM) for 24 hours at 37°C. At the end of the treatment, the cells were collected and pelleted by centrifugation at 1,200× g for 5 minutes. The diluted Cyto-ID® green fluorescent stain solution (250 μL) (Enzo Life Sciences Inc.) was added to each sample and mixed well. Cells were incubated for 30 minutes at 37°C in the dark. After the incubation, cells were washed with 1× assay buffer and resuspended in 250 μL fresh 1× assay buffer. Thereafter, cells were analyzed using the green fluorescence (FL1) channel of the flow cytometer that collected 20,000 events for analysis.

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 - AML12 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 - AML12 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.