Acridine Orange hemi(zinc chloride) salt

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	10μM Acridine Orange to stain the cells and observe by Fluorescence microscope or Flow Cytometry.

Protocol tips
10μM Acridine Orange to stain the cells and observe by Fluorescence microscope or Flow Cytometry.

Publication protocol

Cells (1 × 104 cells/mL) were plated in 60 mm plates and treated with quercetin (0–100 μM) for 48 h. Then, the cells were harvested, washed with phosphate-buffered saline (PBS), fixed in 70% ethanol, rehydrated in 2 mM EDTA-PBS, treated with RNase A (25 ng/mL), and stained with propidium iodide (40 μg/mL) for flow cytometry. The cells were stained with 10 μM acridine orange, harvested, and maintained in 2 mM EDTA-PBS containing 10% FBS to detect autophagy. We followed the manufacturer's protocol for JC-1 mitochondrial membrane detection. In brief, treated cells were trypsinized and washed with 1× assay buffer, stained with JC-1 for 15 min at 37°C in a CO2 incubator, and washed twice with 1× assay buffer at room temperature. All analyses were performed using a FACScaliber flow cytometer (BD Biosciences). Data from 10,000 cells per sample were analyzed with CellQuest software (BD Biosciences). Each experiment was repeated at least three times.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Acridine Orange hemi(zinc chloride) salt below.

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