Cells were plated and treated with RAC1 (2 µm ) or irradiated with 200 mJ cm−2 UVB. After the desired post-treatment time, cells were harvested and stained with APC-Annexin (BD Pharmingen) and propidium iodide (Sigma Chemical). Samples were analyzed by flow cytometry immediately after staining on BD LSR, and the data were analyzed by CELLQuest software.
Apc annexin
Manufactured by BD
Sourced in United States
APC-Annexin is a fluorescent-labeled protein used in flow cytometry applications to detect and quantify apoptosis. It binds to the phospholipid phosphatidylserine, which is exposed on the surface of cells undergoing programmed cell death.
Automatically generated - may contain errors
2 protocols using apc annexin
1
Apoptosis Quantification by Flow Cytometry
2
Apoptosis and Cell Cycle Analysis
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Apoptosis analysis: HCT116 and SW480 cells transfected with sh-NC or sh-NELFCD were trypsinized and resuspended in 1× binding buffer at 2.5×106 cells/mL. Then, 100 µL of this cell suspension was incubated with 5 µL of 7-aminoactinomycin D (BD Pharmingen, San Diego, CA, USA) and 5 µL APC Annexin (BD Pharmingen) for 15 minutes in the dark. The reaction was terminated with the addition of 400 µL 1× binding buffer and analyzed with flow cytometer (BD Pharmingen) using the Flowjo software.
Cell-cycle analysis: The cells were fixed in 70% ethanol, washed twice with PBS, and then labeled with PI-RNase Staining Buffer (BD Pharmingen) for 30 minutes in the dark. Samples were run on a flow cytometer, and the percentages of cells within each phase of the cell cycle were analyzed using Modfit software (Verity Software House, Topsham, ME, USA).
Cell-cycle analysis: The cells were fixed in 70% ethanol, washed twice with PBS, and then labeled with PI-RNase Staining Buffer (BD Pharmingen) for 30 minutes in the dark. Samples were run on a flow cytometer, and the percentages of cells within each phase of the cell cycle were analyzed using Modfit software (Verity Software House, Topsham, ME, USA).
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