CYTO-ID® Autophagy detection kit

Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)

Experiment
Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)
Product
CYTO-ID® Autophagy detection kit from Enzo Life Sciences
Manufacturer
Enzo Life Sciences

Protocol tips

Upstream tips
250 µL of the diluted Cyto-ID® green stain solution was added to each sample and mixed well instead of 100ul
Protocol tips
Use Cyto-ID green dye (1 μl/4 ml assay buffer) for 30 minutes in dark at RT.
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

The cells were treated with fresh medium alone, control vehicle alone (0.05% DMSO, v/v), or ALS at 0.1, 1, and 5 µM for 24 hours at 37°C. To investigate the mechanisms for ALS-induced autophagy, cells were pre-treated with 20 µM Z-VAD(OMe)-FMK, 10 µM WM, 0.1 µM bafilomycin A1, 30 µM chloroquine, 20 µM SB202190, or 25 nM FK866 for 1 hour, then treated with 5 µM ALS for a further 24 hours. Groups of cells treated with each of these compounds alone were also included. All inducers and inhibitors were dissolved in 0.05% DMSO. After ALS treatment, cells were re-suspended in 250 µL of phenol red-free culture medium (Invitrogen Inc.) containing 5% FBS, and 250 µL of the diluted Cyto-ID® green stain solution (Enzo Life Sciences Inc.) was added to each sample and mixed well. Cells were incubated for 30 minutes at 37°C in the dark, collected by centrifugation, washed with 1× assay buffer, and re suspended in 250 µL fresh 1× assay buffer. Cells were finally analyzed using the fluorescein isothiocyanate channel of a flow cytometer that collected 10,000 events for analysis.

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

Reviews

CYTO-ID® Autophagy detection kit from Enzo Life Sciences has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells) using CYTO-ID® Autophagy detection kit from Enzo Life Sciences.

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

Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells) 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.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms