Annexin-V-fluorescein isothiocyanate (FITC) Apoptosis Detection Kit (R&D Systems, Abingdon, UK) was utilized to detect the apoptosis of NSCLC cells. Briefly, NSCLC cells were collected and then re-suspended in 100 μL binding buffer. Annexin-V-FITC and propidium iodide (PI) were incubated with NSCLC cells in the dark for 15 min. The apoptosis of NSCLC cells was analyzed on FC-500 flow cytometer (Beckman Coulter, Pasadena, CA, USA).
Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit
Manufactured by R&D Systems
Sourced in United Kingdom
The Annexin-V-fluorescein isothiocyanate (FITC) Apoptosis Detection Kit is a lab equipment product that allows for the detection and quantification of apoptosis in cell populations. 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. The kit provides Annexin V conjugated to the fluorescent dye FITC, enabling the visualization and analysis of apoptotic cells using flow cytometry or fluorescence microscopy.
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4 protocols using annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit
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Annexin-V-FITC Apoptosis Assay
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Annexin-V-FITC Apoptosis Assay
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Annexin‐V‐fluorescein isothiocyanate (FITC) Apoptosis Detection Kit (R&D Systems, Abingdon, UK) was used in this study to detect the apoptosis of HCC cells. HCC cells were suspended in binding buffer followed by dyeing with propidium iodide (PI) and Annexin‐V‐FITC. After washing using PBS, the samples were analyzed through a flow cytometer (BD Biosciences, San Jose, CA, USA).
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Gemcitabine and ASA Cytotoxicity Assay
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ASA was purchased from Sigma-Aldrich (Merck KGaA) and was dissolved in dimethyl sulfoxide (DMSO; SRL Chemical) to make a 5-M stock solution. This solution was stored at −20°C. Gemcitabine was obtained from Eli Lilly and Company and dissolved in a sterile saline solution to make a 1g/l stock solution. This solution was stored at −20°C. Even though different solvents were used for Gemcitabine and ASA due to their different solubilities, this was not anticipated to significantly influence the results obtained. In addition, the concentration of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; Merck KGaA) was set to 5 mg/ml. The Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit was obtained from R&D Systems, Inc.
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Quantifying Apoptosis with Annexin V-FITC
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The magnitude of apoptosis was detected using an annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit (R&D Systems Inc.) according to manufacturer instructions. In brief, the collected cells were washed with cold PBS, fixed in 75% ethanol at 4°C for 30 min and then stained with annexin V-FITC and PI for 20 min at RT in the dark. Using a flow cytometer, the fluorescence intensities of the cells were quantified as percentages of annexin V-positive and PI-negative (annexin V + /PI -) cell populations, as indicators of apoptotic cells, while the V -/PI -cell population was considered normal (Zhang et al. 2020) (link).
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