Cell apoptosis was assessed by Annexin V-FITC and propidium iodide staining (BD Sciences). Briefly, 9×104 pancreatic cancer cells/well were plated in six-well dishes overnight and treated with different concentrations of CuI for 24 h. The treated cells were washed with PBS, collected, and stained with Annexin V antibody conjugated with a FITC fluorophore and PI in the dark at room temperature. For each measurement, at least 1×104 cells were analysed on an Accuri C6 flow cytometer (Becton Dickinson) using CFlow Plus software.
Fitc annexin 5 and propidium iodide staining
Manufactured by BD
Sourced in United States
FITC-Annexin V and Propidium iodide staining is a laboratory technique used for the detection of apoptosis and cell viability. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is exposed on the outer membrane of apoptotic cells. FITC (Fluorescein isothiocyanate) is a fluorescent dye conjugated to Annexin V, allowing the detection of apoptotic cells. Propidium iodide is a DNA-intercalating dye that can only enter cells with compromised cell membranes, thereby staining necrotic or late-stage apoptotic cells. This combination of staining provides a quantitative assessment of the percentage of viable, apoptotic, and necrotic cells in a sample.
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Lab products found in correlation
Annexin 5 staining kit, by BD (3 mentions)
Cellquest, by BD (3 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Novoexpress software, by Agilent Technologies (1 mentions)
Novocyte flow cytometer system, by Agilent Technologies (1 mentions)
Facsaria 3, by BD (1 mentions)
6 protocols using fitc annexin 5 and propidium iodide staining
1
Quantifying Apoptosis in Pancreatic Cancer
2
Apoptotic Cell-Induced CD8+ T Cell Activation
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For preparing apoptotic cells, splenocytes were incubated with 2.5 μg/mL of cisplatin in DMEM supplemented with 10 % heat-inactivated FBS and 1× of penicillin streptomycin l -glutamine for 24 hours or exposed to UV light for 10 minutes (240 J/s/m2) and incubated for 24 hours in the same medium. Apoptosis was confirmed by annexin V-FITC and propidium iodide staining (BD Pharmingen, 556547) followed by flow cytometric analysis. After washing, cisplatin-induced and UV-induced apoptotic cells (0.5 × 106 cells/well) were cocultured with CD8+ T cells in a 1:1 ratio in 24-well plates. Anti-TIM3 or control IgG was added in some wells, and 48 hours later, intracellular IL-13 was analyzed by flow cytometric analysis.
3
Annexin V Cell Viability Assay
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Cell viability was measured using an annexin V staining kit (BD Biosciences, San Jose, CA, USA). Briefly, cells were treated with the selected alkaloids for 24 h. Cells were then harvested and analysed by multiparametric flow cytometry using FITC-Annexin V and Propidium iodide staining (BD Biosciences, San Jose, CA, USA) according to the manufacturer's instructions. Flow cytometry was then carried out using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA). Data acquisition and analysis was performed with CellQuest (BD Biosciences, San Jose, CA, USA). Data were obtained from three independent experiments.
4
Apoptosis Evaluation of BV-2 Cells
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Cell apoptosis of BV-2 cells was measured by flow cytometry using the annexin V staining kit (BD Biosciences, San Jose, CA, United States). In brief, BV-2 cells were pretreated with 5 μM Aβ(1-42) for 12 h, then followed with an incubation of 0.12–0.48 mg⋅L-1 LSF. After treatment, BV-2 cells were analyzed by the NovoCyte Flow Cytometer Systems (ACEA Biosciences) using FITC-annexin V and propidium iodide staining (BD Biosciences, San Jose, CA, United States) according to the manufacturer’s instructions. Data acquisition and analysis were performed with NovoExpress software (ACEA Biosciences).
5
Annexin V Assay for Cell Viability
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Cell viability and cell death were measured using an annexin V staining kit (BD Biosciences, San Jose, CA, USA). Briefly, cells were treated with 10 μM of hernandezine for 24 h. Cells were then analysed by multiparametric flow cytometry using FITC-Annexin V and Propidium iodide staining (BD Biosciences, San Jose, CA, USA). Flow cytometry was then carried out using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA). Data acquisition and analysis was performed with CellQuest (BD Biosciences, San Jose, CA, USA). Data were obtained from three independent experiments.
6
Quantifying Apoptosis by Flow Cytometry
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Thalidezine-treated cells were harvested and analysed by multiparametric flow cytometry using FITC-Annexin V and propidium iodide staining (BD Biosciences, San Jose, CA, USA) according to the manufacturer instructions. Apoptotic cells were quantitatively counted by a flow cytometer (BD FACSAria III, San Jose, CA, USA). Data acquisition and analysis were performed with CellQuest (BD Biosciences, San Jose, CA, USA) from triple independent experiments.
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