Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining (BD Biosciences) was used to determine chondrocyte apoptosis. Briefly, the cells were collected and washed thrice with ice-cold PBS. Thereafter, they were incubated with 5u Annexin V-FITC added in 300 μl of binding buffer for 25 min and then stained with 5 μl of PI added in 200 μl of binding buffer for 5 min, at room temperature in the dark. Chondrocyte apoptosis was then evaluated within 30 min using the BD Accuri C6 Plus flow cytometer, and the data were analyzed using the FlowJo software.
Annexin 5 fluorescein isothiocyanate fitc propidium iodide staining
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Sourced in United States
Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining is a laboratory technique used to detect and quantify apoptosis, or programmed cell death, in cell populations. Annexin V is a protein that binds to phosphatidylserine, a phospholipid that is exposed on the surface of apoptotic cells. FITC is a fluorescent dye that is conjugated to Annexin V, allowing the identification of apoptotic cells. Propidium iodide is a dye that stains the DNA of cells with compromised cell membranes, typically late-stage apoptotic or necrotic cells.
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7 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide staining
1
Chondrocyte Apoptosis Assessment
2
Evaluating Chondrocyte Apoptosis via Annexin V-FITC/PI
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Annexin V-fluorescein isothiocyanate (FITC)/ propidium iodide (PI) staining (BD Biosciences) was used to evaluate chondrocyte apoptosis. Cells were collected and washed twice with ice-cold PBS. Cells were incubated with u Annexin V-FITC combined with100 μL binding buffer for 25min and then stained with 5 μL PI for 10 min at room temperature in the dark. Chondrocyte apoptosis was analysed using a BD Accuri C6 Plus flow cytometer within 1 h.
3
Apoptosis Analysis in Hepatoma Cells
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Hepatoma cells were trypsinized, washed twice with cold phosphate buffered saline (PBS) and apoptosis rates were determined using flow cytometric analysis with annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's protocol. A total of 500 µl binding buffer, 5 µl Annexin V-FITC and 5 µl PI was added to the cells. Cells were then incubated in the dark at RT for 10 min. Subsequently, the cell apoptosis rate was analyzed using flow cytometry (FACScan, BD Biosciences). All experiments were repeated ≥3 times.
4
Quantifying RSC96 Cell Apoptosis
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RSC96 cell apoptosis was assessed using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining (BD Biosciences). RSC96 cells were digested with EDTA-free trypsin (Gibco; Thermo Fisher Scientific, Inc.). Cells were washed twice with ice-cold PBS and resuspended in 100 μl binding buffer. Cells were incubated with 5 μl Annexin V-FITC and 5 μl PI for 15 min at room temperature in the dark. Apoptosis was analysed using flow cytometry within 30 min of staining.
5
Apoptosis, Cell Cycle, and Oxidative Stress Assay
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According to the manufacturer's instructions, cell apoptosis was assessed using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining (BD Pharmingen, San Diego, CA, USA). Cell cycle was determined with PI (0.05 mg/mL) and RNase A (0.5 mg/mL) staining. Intracellular ROS generation was measured by incubating cells with 10 mmol/L DCF-DA (Sigma, St. Louis, MO, USA) at 37°C for 30 min. ΔΨm was measured by JC-1 dye solution (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) in the dark at 37°C for 20 min. The cells were analyzed using a FACScan flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA).
6
Apoptosis Detection via Annexin V-FITC/PI
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According to the manufacturer’s instructions, apoptosis was determined by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining (BD-PharMingen, San-Jose, CA, USA). Briefly, Capan-2 cells were collected in 6-well plates at a concentration of 105 cells/mL. Then, Annexin V-FITC (5 μL) and PI (5 μL) were distributed into each well and incubated for 15 minutes in the dark for flow cytometry (BD-LSRII, San Jose, California).
7
Apoptosis Detection by Annexin V-FITC/PI
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PIG1 and PIG3V were plated into 6-well plates at a density of 2-3×105 cells/well. Cell apoptosis was detected by using Annexin V−fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining (BD Biosciences, USA). In brief, after disposal by PF and H2O2 like cell cycle, cells were harvested and then washed with cold PBS. After centrifugation, cells were resuspended with Annexin V−FITC (5 µl) and PI (5 µl) for 15 min in the dark. A flow cytometer (BD Biosciences, USA) was used to analyze the samples.
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