Annexin v 7aad staining kit

Mitochondrial membrane potential (ΔΨm) was detected by a fluorescent JC-1 dye. MCF7 cells transfected with vector control and FBXL20 for 36 h were then either treated with DMSO or 5 μm Camptothecin for 12 h. Cells were washed twice with PBS and resuspended in serum-free DMEM medium containing JC-1 dye (Santacruz, sc-364116) for 30 min at 37 °C and analysed by flow cytometry. The fluorescence emission shift from red to green was determined as a change in mitochondrial membrane potential.
Percentage of apoptotic cells was determined using Annexin-V/7AAD staining kit (Biolegend). MCF7 cells expressing NS or FBXL20 shRNA or PUMA shRNA or BAX shRNA were treated with either DMSO or Doxorubicin 5 μM, 16 h. Cells were collected, washed with PBS/0.1% BSA, and incubated with FITC-conjugated Annexin V and 7AAD. Stained cells were analyzed by flow cytometry.

For CD45–20D5, RMA cells
were preincubated with 20 ng/mL of IFNγ for 2 days. For CD45–5E6,
RMA cells did not require IFNγ stimulation. Target cells were
labeled with 1 μM cell trace violet (CTV) (Thermo Fisher) for
20 min at 37 °C. Effector NK cells were co-cultured with 10,000
target cells at different effector/target ratios in the presence of
100 nM CD45-NKR molecules. After 4 h, the cells were spun down, washed
with PBS, and assayed with Annexin V/7-AAD staining kit (BioLegend).
The cells were stained with Annexin V, diluted in 1x Annexin V buffer
for 15 min, and incubated with 2x 7-AAD for 5 min. Dead cells defined
as AnnexinV+ and/or 7AAD+ cells were detected in a CytoFlex flow cytometer
(Beckman Coulter). The cells were stained with Annexin V, diluted
in 1x Annexin V buffer for 15 min, and incubated with 2x 7-AAD for
5 min. Dead cells defined as AnnexinV+ and/or 7AAD+ cells were detected
in a CytoFlex flow cytometer (Beckman Coulter).Dead cells were defined
as Annexin V+. The cells were stained in a Cytoflex cytometer with
Annexin V, diluted in 1x Annexin V buffer for 15 min, and incubated
with 2x 7-AAD for 5 min.