The assessment of apoptotic cells was performed according to previous work [22 (link)]. Briefly, cells were pretreated with LB extracts 2 hr, respectively, prior to UVB irradiation for 24 hr. The changes of early and late apoptosis were determined using an Annexin-V-Fluorescein isothiocyanate (FITC)/PI Apoptosis Detection kit (BioVision, CA 94043, USA). Briefly, cells were resuspended in binding buffer then incubated in the dark with FITC-labeled Annexin V and propidium iodide for 15 min at room temperature; finally, the samples were diluted with phosphate-buffered saline (PBS). Flow cytometry was carried out on a FACScan instrument (FACS Calibur; Becton Dickinson, Mountain View, CA, USA), and data were processed with WinMDI/PC-software V2.9 (written by Joseph Trotter, Scripps Research Institute, La Jolla, CA, USA). Cells labeled with annexin conjugated with the FITC fluorescence were recognized as apoptotic populations.
Annexin 5 fluorescein isothiocyanate fitc pi apoptosis detection kit
Manufactured by Abcam
Sourced in United States
The Annexin-V-Fluorescein isothiocyanate (FITC)/PI Apoptosis Detection kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death, in cell populations. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye, to distinguish between viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometry or fluorescence microscopy.
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2 protocols using annexin 5 fluorescein isothiocyanate fitc pi apoptosis detection kit
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Evaluating Apoptosis Levels via Annexin V-FITC/PI Assay
2
Quantifying Apoptosis by Flow Cytometry
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Apoptosis rates were measured by flow cytometry using the AnnexinV-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (BioVision, USA), as described [25] . After treatment with or without OGD-R, cells were collected at a concentration of 1 × 10 5 cells/mL (total 8 mL for each), mixed with Annexin V-FITC and propidium iodide according to manufacturer's recommendation, and analyzed using a flow cytometer. Data were analyzed using the Cell Quest software (BD Biosciences, USA).
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