PE Annexin V Apoptosis Detection Kit I

Protocol tips

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	Add 5 µl of PE Annexin V and 5 µl 7-AAD

Protocol tips
Add 5 µl of PE Annexin V and 5 µl 7-AAD

Publication protocol

Apoptosis assay

OVCA cells were seeded in 6 well plates (TPP Techno Plastic Products) at a density of 0.25 X105 cells/well and allowed to attach overnight. Different OVCA cells were exposed to 0.5 uM SP-6-27 concentration or with DMSO vehicle control. After 24 or 48 hrs, floating and attached cells were collected, and washed with PBS. Annexin V /7 AAD staining (BD Pharmingen, San Diego, CA, USA) was used to detect apoptosis according to the manufacturer’s instructions. For this, 1 × 105 cells were resuspended in 250 μl of binding buffer and 5 μl of Annexin V-fluorescein isothiocyanate and 5 μl of 7-AAD. Cells were incubated at room temperature for 15 min in dark, after which the cells were washed, resuspended in 300 μl binding buffer and immediately analyzed in flow cytometer (FACSCalibur, Becton Dickinson, Franklin Lakes, NJ, USA). The experiment was performed at least twice in triplicate

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