Annexin V-FITC Apoptosis Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
• Detection Method Flow cytometry (Ex = 488 nm; Em = 530 nm) and fluorescence microscopy • Species Reactivity: Mammalian • Applications: Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.	• Seed 100,000 cells/mL in 6-well plates.

Upstream tips
• Detection Method Flow cytometry (Ex = 488 nm; Em = 530 nm) and fluorescence microscopy • Species Reactivity: Mammalian • Applications: Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.
Protocol tips
• Seed 100,000 cells/mL in 6-well plates.

Publication protocol

Cells were treated with 3.2 mM, 4.8 mM, or 6.4 mM aspirin or left untreated for 24 hours. The cells were then trypsinized with TrypLE express solution and analyzed for apoptotic, necrotic, and viable cell populations using the Annexin V-FITC Apoptosis Kit (BioVision K101-25) according to our previous report.39 Cells were resuspended in 500 μL of 1× Annexin-binding buffer (50 mM HEPES, 700 mM NaCl, 12.5 mM CaCl2, pH 7.4) (Catalog no. V13246, Thermo Fisher Scientific, Mississauga, ON L5N 5V4, Canada) for flow cytometry followed by 5 μL of each of Annexin V-FITC and propidium iodide. The cells were kept at room temperature for 5 minutes in the dark before being quantified by flow cytometry analysis

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