Apoptosis was analyzed by TUNEL assay, and flow cytometry. TUNEL staining (Beyotime Biotechnology, China) was performed following the manufacturer’s instructions. For Annexin V-FITC staining (Beyotime Biotechnology, China), the cells were suspended in 100 μL of Binding Buffer and then were treated with 5 μL Annexin V-FITC and 5 μL of PI in the dark for 15 min. Finally, they were analyzed by flow cytometry.
Annexin 5 fitc staining
Manufactured by Beyotime
Sourced in China
Annexin V-FITC staining is a laboratory technique used to detect and quantify apoptosis, or programmed cell death, in cell samples. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is exposed on the outer cell membrane during apoptosis. The FITC (fluorescein isothiocyanate) label allows the detection of Annexin V using fluorescence-based methods, such as flow cytometry or fluorescence microscopy.
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2 protocols using annexin 5 fitc staining
1
Apoptosis Analysis by TUNEL and Annexin V
2
Doxorubicin and Fulvestrant Induce Apoptosis
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Cells were treated with doxorubicin and fulvestrant alone or in combination for 48 h and the cell morphology was identified using a light microscope at × 400 magnification. Annexin V-FITC staining (Beyotime, Haimen, China) was used to detect cell apoptosis according to the manufacturer's instructions (27 (link)). Briefly, after the designated treatment and at the end of time-point, both detached and attached cells were harvested and washed twice with ice-cold PBS. The collected cells were then resuspended with Annexin V binding buffer and incubated with 5 µl of fluorescein isothiocyanate (FITC) Annexin V for 15 min at 4°C in the dark. The percentages of apoptotic cells were determined by flow cytometry.
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