Acridine Orange hemi(zinc chloride) salt

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Intercalates between DNA/RNA and used to analyze mitochondria and Lysosomal content by flow cytometry.	Stain cells with acridine orange (1 μg/ml) for 15 mins at 37°C

Upstream tips
Intercalates between DNA/RNA and used to analyze mitochondria and Lysosomal content by flow cytometry.
Protocol tips
Stain cells with acridine orange (1 μg/ml) for 15 mins at 37°C

Publication protocol

Cells were treated with DMSO or celecoxib (8 and 30 μM) for 72 hours. In some cases, U87MG cells were pre-treated with PFT (25 μM) for 30 minutes prior to celecoxib treatment. For apoptosis assays, trypsinised cells were incubated with FITC-conjugated Annexin V (BD Biosciences) and propidium iodide (5 μg/ml). 10,000 events were analysed for apoptosis by FACS with CellQuest Pro software. For autophagy assays, celecoxib-treated cells were stained with acridine orange (1 μg/ml, Sigma) for 15 mins at 37°C. Trypsinised cells were re-suspended in phenol red-free growth media (Gibco BRL) and 10,000 events were analysed by FACS with CellQuest Pro software. Acridine orange-stained cells grown on cover slips were viewed under a Laser Scanning Microscope and images captured using software LSM510.

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