To quantify cell death and apoptosis, cells were harvested by digesting with 0.25% trypsin without EDTA after washing in cold PBS. Proteolysis was then neutralized with fetal bovine serum, and the lysates were concentrated and resuspended in 100 μL PBS. Annexin V-FITC and PI staining solution (Beyotime China) were added. After incubated in the solution for 10–20 min at room temperature away from light, the cell suspension was analyzed for apoptosis by flow cytometry.
Annexin 5 fitc and pi staining solution
Manufactured by Beyotime
Sourced in China
Annexin V-FITC and PI staining solution is a laboratory reagent used for the detection and quantification of apoptotic cells. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine (PS), which is externalized to the outer leaflet of the plasma membrane during the early stages of apoptosis. FITC (Fluorescein Isothiocyanate) is conjugated to Annexin V, allowing for the fluorescent labeling of apoptotic cells. Propidium Iodide (PI) is a DNA-binding dye that can penetrate the compromised membranes of late apoptotic or necrotic cells, staining their nuclei. Together, Annexin V-FITC and PI staining solution can be used to distinguish between viable, early apoptotic, late apoptotic, and necrotic cells in a cell population.
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Lab products found in correlation
3 protocols using annexin 5 fitc and pi staining solution
1
Apoptosis Quantification by Flow Cytometry
2
Chondrocyte Apoptosis Measurement
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After transfection 48 h, CHON-001 chondrocytes were digested by trypsin. The chondrocytes were stained by Annexin V-FITC and PI staining solution (Beyotime, Shanghai, China) for 15 min. Flow cytometry was used to measure the chondrocyte apoptosis rate.
3
Quantifying Cell Death in NMCMs
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To quantify cell death, NMCMs were harvested by digesting with 0.05% trypsin without EDTA after washing in cold PBS. Proteolysis was then neutralized with fetal bovine serum, and the cell suspension were concentrated and resuspended in 100 μL PBS. Annexin V-FITC and PI staining solution (Beyotime) were added, and the suspension was subjected to flow cytometry (Backman Coulter, USA, CytoFLEX). Live NMCMs were set on the FSC/SSC dot pattern of unstained control cells ( Supplementary Fig. 8a ) . And the singlet population were further confirmed by FSC-H/FSC-A dot pattern of unstained control cells ( Supplementary Fig. 8b ) . Three cell subpopulations identified from the singlet population of Annexin V-FITC & PI stained A/R cells with unstained control cells as reference ( Supplementary Fig. 8c, d ) .
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