Single-cell suspensions of fetal and postnatal ENCCs at day 5 were collected by centrifugation to assess cell apoptosis by flow cytometry using an Annexin V-fluorescein isothiocyanate/propidium iodide Apoptosis Detection Kit (Cat# 40302ES20, Yeasen, Shanghai, China) as described in our previous studies (Yu et al., 2015). Briefly, 1 × 105 cells suspended in 500 μL of binding buffer were mixed with 5 μL Annexin V-fluorescein isothiocyanate and 5 μL propidium iodide and incubated for 10 minutes at room temperature. Apoptosis was measured with FACSCalibur and CELLQuest software version 2.0 (Becton Dickinson) within 1 hour according to the manufacturer’s instructions.
Annexin 5 fluorescein isothiocyanate propidium iodide apoptosis detection kit
Manufactured by Yeasen
Sourced in China
The Annexin V-fluorescein isothiocyanate/propidium iodide Apoptosis Detection Kit is a laboratory assay used to detect and quantify apoptosis, a form of programmed cell death. The kit includes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a dye that stains DNA. Together, these components enable the identification of cells undergoing early and late stages of apoptosis.
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2 protocols using annexin 5 fluorescein isothiocyanate propidium iodide apoptosis detection kit
1
Apoptosis Assessment of Fetal and Postnatal ENCCs
2
HESC Apoptosis Detection by Flow Cytometry
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The apoptosis level of HESCs was detected using the annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit (40302ES60; YEASEN, China). After drug treatment for 48 hours, or knockdown of ITPR1 for 24 hours, cells were washed 3 times with PBS and harvested into centrifuge tubes. Then, the cells were stained with annexin V/propidium iodide according to the manufacturer's protocol. Finally, the fluorescence-activated cell sorting Calibur flow cytometer (BD Biosciences) was used to detect the apoptosis rate of the HESCs.
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