Sub‐confluent monolayered NHEK cultures in 6‐cm dishes were stimulated with or without IL‐4 + IL‐13 (50 ng/mL) for 48 hours. Then, cells were washed with cold PBS and stained with Annexin V Phycoerythrin (PE) Apoptosis Detection Kit I (BD Biosciences) according to the manufacturer's instructions. Dead cells, which are positive for both 7‐amino‐actinomycin D (7‐AAD) and Annexin V‐PE, were quantified by flow cytometry using a FACS Canto II (BD Biosciences).
Annexin 5 phycoerythrin pe apoptosis detection kit 1
Manufactured by BD
The Annexin V Phycoerythrin (PE) Apoptosis Detection Kit is a laboratory assay used to detect and quantify apoptosis, a programmed cell death process. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, a molecule that is exposed on the surface of cells undergoing apoptosis. Annexin V is conjugated with the fluorescent dye phycoerythrin (PE) to enable detection and quantification of apoptotic cells using flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur flow cytometer, by BD (2 mentions)
Facscanto 2, by BD (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Pe conjugated anti human cd24 antibody, by BD (1 mentions)
Cellquest pro software, by BD (1 mentions)
4 protocols using annexin 5 phycoerythrin pe apoptosis detection kit 1
1
Apoptosis Quantification in NHEK Cells
2
Cell Cycle and Apoptosis Analysis
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A propidium iodide cell cycle detection kit (Biotechnology's Beyotime Institute, Beijing, China) was used to analyze the cell cycle on a FACSCalibur flow cytometer (BD Biosciences, NY, USA). An Annexin V-phycoerythrin (PE) Apoptosis Detection kit I (BD) was used to perform the apoptosis assay, according to the manufacturer instructions. The results were also analyzed using a FACSCalibur flow cytometer.
3
Antileishmanial Activity Evaluation Protocol
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Benzoxonium chloride was purchased from BOC Sciences, USA. Meglumine antimoniate (MA, Glucantime®) 1.5 g/5 mL solution was obtained from Sanofi-Aventis, France. MTT powder (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide; Thiazolyl blue), sorbitan esters (Span®), their PEGylated derivatives (Tween®) and cholesterol (CHOL) were provided by Sigma-Aldrich® (USA). DMEM medium with sodium pyruvate, RPMI- 1640 medium with stable glutamine were purchased from Biosera (France) and fetal bovine serum (FBS) were prepared by Gibco (USA). Phycoerythrin (PE) Annexin V Apoptosis Detection Kit I (catalog number: 559763) was prepared by BD Pharmingen™. Phosphate buffered saline (PBS) was purchased from Cassion Lab (USA). Penicillin and streptomycin were obtained from Life Technology (USA) and was frozen at − 20 °C until testing.
4
CD24 Expression and Apoptosis Profiling
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For CD24 staining, cells (1 × 106/100 μL stain buffer) were incubated with PE-conjugated anti-human CD24 antibody (cat #, 560991; BD Biosciences) for 30 min at 4 °C in dark. PE-conjugated IgG2a, κ-Isotype (cat #, 555574; BD Biosciences) was used as a control. Data were acquired on a BD FACSCalibur flow cytometer (BD Biosciences) using BD CellQuest Pro software (BD Biosciences).
For the apoptosis assay, the spheroid cells were exposed to CDDP (5 μM) for 72 h under serum-free medium and stained with PE Annexin V and 7-Amino-Actinomycin D (7-AAD) for discrimination of early and late apoptosis, respectively, using the Phycoerythrin (PE) Annexin V Apoptosis Detection Kit I (BD Biosciences).
For the apoptosis assay, the spheroid cells were exposed to CDDP (5 μM) for 72 h under serum-free medium and stained with PE Annexin V and 7-Amino-Actinomycin D (7-AAD) for discrimination of early and late apoptosis, respectively, using the Phycoerythrin (PE) Annexin V Apoptosis Detection Kit I (BD Biosciences).
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