The flow cytometry was used to study cell apoptosis. Briefly, cells treated or untreated were dyed using annexin V-fluorescein (BD Pharmingen, Pasig City, Philippines) and propidium iodide (PI) (BD). Samples were then analyzed with BD FACSCanto flow cytometer to determine percentages of apoptotic cells (including early apoptotic cells).
Annexin 5 fluorescein
Manufactured by BD
Sourced in United States
Annexin V-fluorescein is a protein that binds to phosphatidylserine, a component of the cell membrane. It is commonly used in cell biology research to detect and quantify apoptosis, or programmed cell death, in various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Pi apoptosis detection kit, by BD (5 mentions)
Fitc propidium iodide apoptosis detection kit, by BD (3 mentions)
Cellquest 3, by BD (2 mentions)
Facscanto system, by BD (2 mentions)
Facscan flow cytometer, by BD (2 mentions)
Facscanto, by BD (1 mentions)
Facs flow cytometry, by BD (1 mentions)
Moflo xdp flow cytometer, by Beckman Coulter (1 mentions)
Lsr 2 flow cytometry kit, by BD (1 mentions)
Facscan flow cytometry, by BD (1 mentions)
Moflo xdp, by Beckman Coulter (1 mentions)
Macsquant, by Miltenyi Biotec (1 mentions)
Facscanto 2, by BD (1 mentions)
Facs cytometry, by BD (1 mentions)
Flow cytometry, by BD (1 mentions)
Facscan, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cell cycle staining buffer, by MultiSciences Biotech (1 mentions)
Facscanto flow cytometer, by BD (1 mentions)
21 protocols using annexin 5 fluorescein
1
Quantifying Cell Apoptosis by Flow Cytometry
2
Quantitative Analysis of Compound 3a-Induced Cell Death
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The quantitative analysis of compound 3a -induced cell death was performed using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (BD Biosciences, BD Pharmingen™, San Jose, CA, USA) as previously described [32 (link)]. Briefly, the RPMI 8226 or A549 cells growing on the 6-well plates in the culture medium with 2% FBS were subjected to increasing concentrations of compound 3a (2.5, 5, 10, 20, or 40 μM) for 24 and 48 h. After the treatment, the cells were harvested, centrifuged at 1000 rpm and room temperature (RT) for 5 min, washed twice with PBS, and stained with 5 mM of AnnexinV-FITC and 5 mM of PI. After incubation for 15 min in the dark at room temperature, the cells were immediately analyzed using FACS Calibur. The intensity of fluorescence of AnnexinV-FITC and PI stained cells was determined by acquisition of at least 10,000 events per sample within the rate >60 events/s. The FACS data were analyzed using Cell Quest Pro Version 6.0. for the Macintosh operating system. The data were analyzed to determine the percentage of viable, early apoptotic, late apoptotic, and necrotic cells.
3
Apoptosis Assay in OS Cells
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OS cells were seeded in 6-cm plates and incubated with different concentrations of AIL. After 24 h, all the cells including the floating and adherent ones were harvested and stained with Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) apoptosis detection kit (#556547, BD Biosciences Pharmingen, San Diego, CA, USA) according to the manufacturer’s protocols. The resulting fluorescence was assessed by fluorescence-activated cell sorting scan (FACS) flow cytometry (Becton Dickinson, Mountain View, CA, USA) as reported previously (25 (link)).
4
Apoptosis Quantification in PC Cells
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GSP-induced apoptosis in PC cells was determined by flow cytometry using an Annexin V-fluorescein isothiocyanate (FITC) Apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA). Following treatment with GSPs for 48 h at 37°C, cells (2×105) were harvested, washed twice with PBS and incubated with Annexin V-FITC and propidium iodide for 10 min in the dark at room temperature. The stained cells were then detected and analyzed by a MoFLO XDP flow cytometer (Beckman Coulter, Inc., Brea, CA, USA) and the Cell Quest 3.3 software (BD Biosciences).
5
Apoptotic Effects of MT on 4T1 Cells
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4T1 cells were treated with MT (0.2, 0.4, 0.8 mM) for 48 h, harvested and the numbers of cells counted. Apoptosis was evaluated by Annexin V-fluorescein isothiocyanate/propidium iodide (PI), according to manufacturer's instructions (BD Pharmingen; BD Biosciences, Franklin Lakes, NJ, USA) and analyzed using a FACScan flow cytometer (BD FACSDiva software version 8.0.1; BD Biosciences).
6
Rosiglitazone-Induced Apoptosis Pathway
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EC109 and TE10 cells were treated with 20 μM RGZ for 48 h, and apoptosis rate was measured using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (BD PharMingen, San Diego, CA, USA). Cells were collected, and then washed twice with cold PBS and resuspended in 500 μl of Annexin V-FITC binding buffer. The suspension was incubated with 5 μl of Annexin V-FITC and 5 μl of PI at room temperature in the dark for 10 min. Finally, the apoptosis rate was analyzed by fluorescence-activated cell sorting using a BD LSR II flow cytometry kit (BD PharMingen). The apoptosis rates of cells transfected with specific siRNAs targeting RXRα, PPARg, TLR4, MyD88, ERK, JNK, and p38 or control siRNA were measured using FCM analysis described above. Each experiment was conducted for three times independently.
7
Quantifying Neuronal Apoptosis by Flow Cytometry
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Flow cytometry was performed to examine neuronal cell apoptosis using the Annexin V-fluorescein isothiocyanate (AnV-FITC) and propidium iodide (PI) kit (BD PharMingen, Santiago, CA, United States) (Yu et al., 2015 (link)). After OGD/RP treatment, 1 × 106 cells were collected from each sample and seeded into 24-well plates. Cells were incubated with AnV-FITC at room temperature for 15 min, followed by the addition of PI. After 30 min, cells were ready for flow cytometric analysis, and the rate of apoptosis was determined.
8
Apoptosis Assessment of ox-LDL-Induced HAECs
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The apoptosis rate of ox-LDL-induced HAECs was assessed through the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (BD Biosciences, San Jose, CA, USA). Briefly, ox-LDL-induced HAECs with or without transfection were cultured in the medium for 48 h. After washing, the cells were re-suspended in binding buffer (200 µl) at a concentration of 2 × 105 cells/ml. Following this, the cells were stained with Annexin V-FITC (5 µl) and PI (100 µl) for 30 min in the dark. Eventually, the apoptosis rate of ox-LDL-induced HAECs with or without transfection was assessed through the FACScan flow cytometry (BD Biosciences).
9
Apoptosis Quantification by Flow Cytometry
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The cells were cultured in suspension for 48 h and digested with trypsin without ethylenediaminetetraacetic acid (EDTA). Next, flow cytometry analysis was performed using the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (BD Biosciences) according to the manufacturer's instructions. Data were collected on a BD FACS Canto system and analyzed using the Flow Jo software.
10
Flow Cytometric Apoptosis Assay of GSP-Treated Cells
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GSP-induced apoptosis in PC cells was determined by flow cytometry using the Annexin V-fluorescein isothiocyanate (FITC) Apoptosis Detection kit (BD Biosciences, Franklin Lakes, NJ, USA). Following treatment with GSPs for 48 h, 2×105 cells were harvested, washed twice with PBS and incubated with Annexin V-FITC and propidium iodide for 10 min in the dark at room temperature. The stained cells were then detected and analyzed by the MoFlo XDP flow cytometer (Beckman Coulter, Inc., Brea, CA, USA) and Cell Quest 3.3 software (BD Biosciences, Franklin Lakes, NJ, USA).
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