CYTO-ID® Autophagy detection kit

Autophagy assay cell type - Human fetal osteoblastic (hFOB) 1.19

Experiment
Autophagy assay cell type - Human fetal osteoblastic (hFOB) 1.19
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
Incubate cells with CYTO-ID dye (2μl of CYTO-ID in 1ml of 1X assay buffer) for 30 min
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 hFOB 1.19 cells (1 × 104) were seeded on a Lab‐Tek™ II Chamber Slide (Nunc; Thermo Fisher Scientific, Rochester, NY) for 24 h. The next day, the cells were treated with galangin (5 µM), autophagy inhibitor 3‐MA (500 µM), and autophagy inducer rapamycin (50 nM) for 24 h. CYTO‐ID® Green autophagy dye was used to detect autophagy in live cells.26 The autophagosomes and autolysosomes were stained using a CYTO‐ID® Autophagy Detection Kit (Enzo Life Sciences, Farmingdale, NY) and analyzed using confocal microscopy (Carl Zeiss, Germany). Staining was conducted following manufacturer’ protocol

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - Human fetal osteoblastic (hFOB) 1.19 using CYTO-ID® Autophagy detection kit from Enzo Life Sciences.

Paper title
Cytoprotective effect of flavonoid‐induced autophagy on bisphosphonate mediated cell death in osteoblast
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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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