Cells were seeded in triplicate into 24-well plates at a concentration of 2.0 × 105 cells/well, and then treated with 3 different concentrations of M802 or Herceptin. After 48 h at 37 °C and 5% CO2, cells were dissociated with trypsin/EDTA (Gibco), washed twice with cold PBS, and then labeled with Annexin V/PI (BioLegend) as described previously [32 (link)].
Annexin 5 pi
Manufactured by BioLegend
Sourced in United States
Annexin V/PI is a fluorescent labeling solution used to detect and analyze apoptosis, or programmed cell death, in cell populations. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is externalized during the early stages of apoptosis. Propidium iodide (PI) is a fluorescent dye that binds to the DNA of cells with compromised cell membranes, indicating late-stage apoptosis or necrosis. This combination allows for the detection and differentiation of viable, early apoptotic, and late apoptotic/necrotic cells using flow cytometry or fluorescence microscopy.
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Lab products found in correlation
3 protocols using annexin 5 pi
1
Cytotoxicity Assay for M802 and Herceptin
2
Annexin V Apoptosis Assay
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To assess apoptosis, 5 × 105 KG-1α or Kasumi-1 cells were inoculated in 24-well plates containing 1 mL medium and treated with designated drugs for 24 h, followed by staining with Annexin V/PI (Biolegend) in dark for 15 min. Then, flow cytometry was used to analyze the percentage of apoptotic cells (Annexin V+).
3
Evaluation of HUVEC Viability and Phenotype
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The viability of HUVEC was assessed by flow cytometry with the determination of the number of viable cells, as well as those in early and late apoptosis and necrosis evaluated by Annexin-V/PI (BioLegend, San Diego, CA, USA) double staining.
The expression of von Willebrand factor and CD146 (BioLegend, San Diego, CA, USA) in HUVECs was detected by immunocytochemical staining. Cell nucleuses were stained with 4′,6-diamidino-2-phenylindole (DAPI).
The expression of von Willebrand factor and CD146 (BioLegend, San Diego, CA, USA) in HUVECs was detected by immunocytochemical staining. Cell nucleuses were stained with 4′,6-diamidino-2-phenylindole (DAPI).
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