FITC Annexin V Apoptosis Detection Kit I (RUO)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 5 µl of FITC Annexin V and 5 µl PI and incubate for 15 min at RT (25°C) in the dark.

Protocol tips
Add 5 µl of FITC Annexin V and 5 µl PI and incubate for 15 min at RT (25°C) in the dark.

Publication protocol

For cell division, 2 × 107 T cells in PBS were labeled with 2.5μM CFSE for 5 minutes at room temperature. Labeled cells were cocultured with tumor cells at 2:1 ratio in the absence of exogenous IL-2. After 5 days, cells were stained with CD3 and analyzed for CFSE dilution by FACS analysis. For expansion, viable T cells were cocultured with tumor cells at 1:1 ratio in the absence of IL-2, and T-cell counts were determined by the Multisizer 3 Coulter Counter (Beckman Coulter) and Trypan blue exclusion on day 5 and 10. For apoptosis assays, T cells were cocultured with tumor cells at 2:1 ratio in the absence of IL-2. After 3 days, T cells were detected by simultaneous staining with anti-CD3 antibody, annexin V–FITC, and 7-AAD-PerCP-Cy5.5, according to the manufacturer's instructions (BD Pharmingen).

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Manufacturer protocol

Download the product protocol from BD Biosciences for FITC Annexin V Apoptosis Detection Kit I (RUO) below.

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