CNE1 cells were seeded onto six-well plates at the density of 10 × 104 cells per well and irradiated with various doses of IR. The cells were then incubated for 48 h, and stained with FITC-conjugated Annexin V and propidium iodide (PI), using Annexin V-FITC Apoptosis Detection kit (Beyotime Institute of Biotechnology, Nantong, China) and according to manufacturer's recommendation. Each sample was then subjected to analyses by flow cytometry using a FACSVantage SE system (BD Biosciences, San Jose, CA, USA). The percentage of apoptotic cells was determined by Flow cytometry.
Fitc conjugated annexin 5
Manufactured by Beyotime
Sourced in China
FITC-conjugated Annexin V is a fluorescently labeled protein that binds to phosphatidylserine, which is exposed on the surface of cells undergoing apoptosis. This property allows it to be used as a marker for the detection of apoptotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Beyotime (4 mentions)
Facsvantage se system, by BD (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (1 mentions)
Fluorescence activated cell sorting (facs), by Beckman Coulter (1 mentions)
Rnase a, by Beyotime (1 mentions)
Flow cytometry, by Beyotime (1 mentions)
Expo32 adc software, by Beckman Coulter (1 mentions)
8 protocols using fitc conjugated annexin 5
1
Apoptosis Assay for IR-Induced Cell Death
2
Apoptosis Assessment via Flow Cytometry
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Propidium iodide (PI) and FITC‐conjugated Annexin V staining (Beyotime) were used for flow cytometry. 1× PBS washed and PI/FITC‐Annexin V–stained cells in 50 μg/mL RNase A (Beyotime). After that, the cells were hatched (1 hour, dark, 25°C). Results were performed through fluorescence‐activated cell sorting (Beckman Coulter). Results were analysed by FlowJo software (Tree Star Software).
3
Apoptosis Measurement by Annexin V-FITC
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Cells were cultured overnight in 6-well plates and treated for 24 h with BSE. Apoptosis in the cultures was assessed using a kit based on staining with FITC-conjugated annexin V (catalog no. C1062S, Beyotime, Shanghai, China) and flow cytometry (Beckman Coulter, Brea, CA, USA). Cells were suspended in a mixture of 1X annexin V-FITC binding buffer (195 µL) and annexin V-FITC (5 µL), incubated at room temperature for 10 min, and centrifuged at 1,000 ×g for 5 min. The pellet was resuspended in binding buffer (190 µL), and propidium iodide (PI) working solution (10 µL) was added before the samples were analyzed using flow cytometry.
4
Annexin V Apoptosis Assay Protocol
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Cell apoptosis was determined using an Annexin V assay as described previously (21 (link)). After transfection and/or lidocaine treatment, cells were collected, washed, and suspended in Annexin V-binding buffer. FITC-conjugated Annexin V and propidium iodide (PI; Beyotime, Haimen, China) were added to cells successively. After incubation, Annexin V-binding buffer was added, and cells were analyzed using a FAC Scan Flow Cytometer.
5
Annexin V/PI Apoptosis Assay
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Cells (1×103 cells/ml) were harvested and washed, then stained with FITC-conjugated Annexin V (Beyotime Institute of Biotechnology) and PI (Beyotime Biotechnology) at room temperature in the dark for 15 min. Cell apoptosis was measured on a COULTER FC500 MCL flow cytometer (Beckman Coulter, Inc.) using EXPO32 ADC software (Beckman Coulter, Inc.). Apoptotic index was calculated by adding the proportion of Annexin V(+)/PI(−) early apoptotic cells to that of Annexin V(+)/PI(+) late apoptotic cells. The experiment was repeated three times with five replicates.
6
Apoptosis and Necrosis Analysis of Transfected 293T Cells
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293 T cells were first transfected with pcDNA3.1-Lnc45 or pcDNA3.1 for 24 h, and then the cells were then inoculated with CK10 virus. At 24 h p.i, the cells were then collected and analyzed as described previously [25 (link)]. Briefly, a total of 106.0 cells was stained with fluorescein isothiocyanate (FITC)-conjugated annexin V and/or propidium iodide (PI) (Beyotime, Shanghai, China), respectively, and were then analyzed using the flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). The ratios of apoptotic or necrotic cells were presented as the means ± standard deviations (SD) from three independent experiments.
7
Apoptosis Assay with Annexin V
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Cell apoptosis was determined using an Annexin V assay and described previously [21] . After transfection and/or lidocaine treatment, cells were collected, washed, and suspended in Annexin V-binding buffer. FITC-conjugated Annexin V and propidium iodide (PI; Beyotime, Haimen, China) were added to cells successively. After incubation, Annexin V-binding buffer was added, and cells were analyzed using a FAC Scan Flow Cytometer.
8
Apoptosis Assay with Annexin V
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Cell apoptosis was determined using an Annexin V assay and described previously [21] . After transfection and/or lidocaine treatment, cells were collected, washed, and suspended in Annexin V-binding buffer. FITC-conjugated Annexin V and propidium iodide (PI; Beyotime, Haimen, China) were added to cells successively. After incubation, Annexin V-binding buffer was added, and cells were analyzed using a FAC Scan Flow Cytometer.
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