The cells were seeded into 6-well plates. After adherence, cells were incubated with drugs for 24 h. The cells were then stained with eBioscienceTM Annexin V Apoptosis Detection Kit (Thermo Fisher Scientific, 88-8007). Analysis was performed using flow cytometry within 4 h.
Ebioscience annexin 5 apoptosis detection kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EBioscience™ Annexin V Apoptosis Detection Kit is a laboratory tool used to detect and measure apoptosis, a programmed cell death process. It provides a simple and effective method for quantifying apoptotic cells based on the translocation of phosphatidylserine from the inner to the outer cell membrane, which is a characteristic of the apoptotic process.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Ma900 cell sorter, by Sony (2 mentions)
Lsrfortessa, by BD (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Fortessa cytometer, by BD (2 mentions)
Canto 2 flow cytometer, by BD (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Tunel detection kit, by Thermo Fisher Scientific (1 mentions)
Facsaria, by BD (1 mentions)
Caspase glo 3 7 assay kit, by Promega (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions)
Mitochondrial membrane potential assay kit, by Beyotime (1 mentions)
Sodium selenite na2seo3, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Cell cycle analysis kit, by Beyotime (1 mentions)
Accuri c6 flow cytometry, by BD (1 mentions)
Fxcycle pi rnase staining kit, by Thermo Fisher Scientific (1 mentions)
26 protocols using ebioscience annexin 5 apoptosis detection kit
1
Apoptosis Analysis via Flow Cytometry
2
Lenvatinib-Induced Apoptosis Analysis
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Cells were seeded in 6-well plates. After adherence, the cells were treated with 10 μM lenvatinib for 24 h. The cells were digested with EDTA-free trypsin and stained using an eBioscienceTM Annexin V Apoptosis Detection Kit (#88-8007, Thermo Scientific) according to the manufacturer’s instructions. Then, Flow cytometry analysis was performed within 4 h.
3
Evaluating Apoptosis and Cell Cycle
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Cell cycle was evaluated as previously reported [39 (link)]. Apoptosis assay was performed after 72 h treatment of emavusertib alone or in combination with ibrutinib by using eBioscience Annexin V Apoptosis Detection Kit (Thermo Fisher Scientific, Basel, Switzerland). Apoptosis induction and cell cycle distribution after the treatment were evaluated with flowCore R package from Bioconductor.
4
T Cell Apoptosis Induction
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CTRL and NSM KD T cells were left unstimulated or stimulated for 24 h with α-CD3 alone or with CD3/CD28-specific Abs in combination as described above (antibody concentration: each 1 µg/ml). When indicated 400 ng/ml FasL (kindly provided by Harald Wajant, University of Würzburg) was added to α-CD3/CD28 stimulated cells for 3 h. Apoptotic cells were detected by flow cytometry using the eBioscience Annexin V Apoptosis Detection Kit (ThermoFisher Scientific).
5
Annexin V-FITC and PI Staining for Cell Death
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Cell death has been evaluated by staining the cells with eBioscience™ Annexin V Apoptosis detection kit (Thermo Fisher Scientific) according to the manufacturer's instructions. Briefly, all cells were harvested using Accutase, washed once with PBS, and once - with 1X Annexin V buffer. 150 000 cells were stained in 200 μL of 1X Annexin V buffer for 15 min at room temperature with FITC-conjugated Annexin V, washed once with 1X Annexin V buffer and re-suspended in 200 μL Annexin V buffer. 5 μL of propidium iodide, PI (Thermo Fisher Scientific) was added to the cell suspension, and cell fluorescence was measured on flow cytometer Canto II (BD Biosciences). Analysis of flow cytometry data was performed using FlowJo software (version 10.6.2).
6
Quantification of Apoptosis in MM Cells
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Apoptosis was detected using an eBioscience™ Annexin V Apoptosis Detection Kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the instructions from the manufacturer. Briefly, MM cells were treated with JA for specific times, and approximately 1 × 106 cells were then harvested and washed once with 1× binding buffer and then resuspended in 100 μL of 1× loading buffer. Then, 5 μL propidium iodide (PI) and 5 μL Annexin V-APC were added per sample to the cell suspension, followed by incubation in the dark for 15 min. The apoptotic cells were quantified using a flow cytometer (Fortessa, San Francisco, CA, USA) using DIVA software. Approximately 10,000 cells were analyzed for each sample.
7
Apoptosis Assay in PDAC Cells
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PDAC cells were treated with Arsenic Trioxide at indicated time points. Samples were tested for apoptosis using an eBioscience Annexin V Apoptosis Detection Kit according to manufacturer instructions (ThermoFisher, Waltham, MA, USA).
8
Quantifying Apoptosis via Annexin V-FITC and PI
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In this assay, levels of the FITC-labeled Annexin V protein indicate apoptosis as the A.V protein binds with high affinity to the phosphatidylserine that is translocated from the inner side of the cell membrane to the outer side. Likewise, levels of propidium iodide (P.I.), which fluoresces upon binding DNA in cells that have ruptured or become permeable, indicate cell death or cells that are in the latest stages of apoptosis(55 (link), 56 ). The preparation of cells for flow cytometry was conducted according to established protocols(55 (link)). Briefly, following the completion of PM exposures using n=3 replicates, culture media was collected and put on ice to recover detached cells. Adherent cells were trypsinized and combined with the collected culture media. The combined cells were washed twice with cold PBS before proceeding with Annexin V-FITC and propidium iodide staining of 250,000 cells per sample using an eBioscience™ Annexin V Apoptosis Detection Kit (ThermoFisher, 88–8005-72). Prepared samples were analyzed on a Sony Biotechnology MA900 Cell Sorter available through the Center for Biomedical Research Support at UT Austin. At least 10,000 cells per replicate were analyzed for Annexin V binding and propidium iodide incorporation.
9
Apoptosis Analysis via Annexin V-FITC
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eBioscience™ Annexin V Apoptosis Detection Kit (cat no. 88-8102-72, ThermoFisher Scientific, USA) was used for analysis of apoptosis according to the manufacturer's instructions. After indicated treatment, the cells were trypsinized, collected, and resuspended. About 2×105 cells were harvested and washed twice with cold phosphate buffer saline (PBS), then resuspended in 500 μl binding buffer. 10μl Annexin V-FITC and 10 μl Propidium Iodide were added to the solution and mixed well. After 15 min incubation, the cells were analyzed using flow cytometric analysis (BD Biosciences, San Jose, CA).
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Annexin V Apoptosis Assay Protocol
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