Apoptotic cells were measured using an Annexin V-fluorescein isothiocyanate (FITC) Apoptosis Detection kit (Nanjing KeyGen Biotech Co., Ltd, Nanjing, China). The cell samples were harvested using 0.25% EDTA-free trypsin after 48 h of incubation, washed twice with ice-cold PBS and resuspended in 500 µl binding buffer. Cells were incubated with 5 µl annexin V-FITC-labeled specific antibodies (from the apoptosis detection kit) and 5 µl propidium iodide (PI) for between 15 and 20 min in the dark, and detected using a FACSAria flow cytometer and FASCDiva 4.1 software (BD Biosciences, Franklin Lakes, NJ, USA). Each experiment was performed in triplicate.
Fascdiva 4
Manufactured by BD
Sourced in United States
The FASCDiva 4.1 software is a data acquisition and analysis tool designed for flow cytometry applications. It provides the core functionality to capture, visualize, and analyze data generated by flow cytometry instruments.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fascdiva 4
1
Quantifying Apoptosis using Annexin V-FITC
2
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis was detected using Annexin V-FITC/PI detection kits (Nanjing KeyGen Biotech Co., Ltd., Nanjing, Jiangsu, China). After 48-h of culture, neurons were collected with 0.25% ethylene diamine tetraacetic acid-free trypsin, washed 2 times with ice-cold PBS, and suspended in 500 μL binding buffer. The cells were then incubated with 5 μL Annexin V-FITC-labeled specific antibody and PI in the dark for 15-20 min, followed by detection in a flow cytometer and analyzed using the FASCDiva 4.1 software (BD Biosciences, Franklin Lakes, NJ, USA). The cells labeled with Annexin V were regarded early apoptotic cells, while cells labeled with Annexin V and PI were regarded as late apoptotic cells. Single positive cells labeled with PI were necrotic cells, while live cells were labeled with neither. The early and late apoptotic cells were counted.
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