CYTO-ID® Autophagy detection kit

Autophagy assay cell type - Fibroblasts

Experiment
Autophagy assay cell type - Fibroblasts
Product
CYTO-ID® Autophagy detection kit from Enzo Life Sciences
Manufacturer
Enzo Life Sciences

Protocol tips

Upstream tips
This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry
Protocol tips
- 2 µL of CYTO-ID® Green Detection Reagent and 1 µL of Hoechst 33342 Nuclear Stain for every 1 mL of 1X Assay Buffer or complete cell growth medium
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 digested and washed twice with PBS (pH 7.4; HyClone, Logan, UT, USA), centrifuged for 5 min at 37°C, and resuspended in a binding buffer. Autophagosomes were marked using the Cyto-ID Autophagy Detection kit (Enzo Life Sciences, Inc., Farmingdale, NY, USA). Marked cells were photographed under a fluorescence microscope (Nikon), and the fluorescence intensity was detected using a flow cytometer (FC500 MPL; Beckman Coulter, Inc., Brea, CA, USA).

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Papers

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

Paper title
TERT alleviates irradiation-induced late rectal injury by reducing hypoxia-induced ROS levels through the activation of NF-κB and autophagy
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

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