CYTO-ID® Autophagy detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry		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
This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry
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

A cell cycle analysis was performed using a Becton Dickinson FACS flow cytometer as previously described [18]. Apoptosis of leukemia cells was assessed based on 7AAD/annexin V staining (Invitrogen, Tokyo, Japan) in cells cultured in the presence or absence of benfotiamine [15] and Giemsa staining was performed [17] as previously described. In some experiments, HL-60 cells were pre-treated with 0.25 μg/ml of either cycloheximide or tetrathylammonium, a big potassium channel (BK) blocker, for 2 hours and then cultured for 12 hours with or without benfotiamine (100 μM) and stained with Giemsa. Autophagy was assessed by Western blotting for LC3 protein or by using the Cyto-ID Autophagy detection kit (Enzo Life Sciences), which uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry.

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 - HL-60 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 - HL-60 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.