Annexin V-FITC Apoptosis Assay Kit
Detect apoptosis and membrane permeability with the Annexin V-FITC Apoptosis Assay Kit. This in vitro apoptosis assay employs the green fluorescent Annexin V-FITC reagent to label exposed phosphatidylserine in cultured cell samples. Detect membrane compromised cells, a trademark of late apoptosis or cell necrosis, with the live/dead stain, propidium iodide. Analyze the fluorescent signals by flow cytometry.
Annexin V-FITC Apoptosis Assay Kit
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- Prepare samples and controls
- Dilute 10X Binding Buffer 1:10 with diH2O.
- Wash cells in ice-cold culture medium or PBS and resuspend in ice-cold 1X Binding Buffer.
- Dilute Annexin V-FITC 1:10 in PBS.
- Dilute Propidium Iodide (PI) 1:2 in PBS.
- Add diluted Annexin V-FITC to each sample at a ratio of 1:20 (5 µL per 100 µL aliquot of cultured cells).
- Add diluted PI to each sample at a ratio of 1:20 (5 µL per 100 µL aliquot of cultured cells).
- Incubate ~10 minutes, protected from light.
- Dilute samples by adding 250 µL 1X Binding Buffer to each.
- Analyze with a flow cytometer. Annexin V-FITC optimally excites at 494 nm and emits at 519 nm (typically read in FL1). PI optimally excites at 536 nm and has a peak emission at 617 nm (typically read in FL3).