Glioblastoma cells were seeded in 6-well plates at 4 × 105 cells/well. The cells were harvested and washed with PBS after being treated for 24 h. Then, the cells were collected and fixed in ice-cold 70% alcohol. After fixation, RNase A (Thermo Fisher Scientific) was added to remove the RNA, and the cells were stained with PI (Sigma) for cell cycle distribution analysis. An APC Annexin V detection kit with 7-AAD (BioLegend) was used to identify apoptotic and necrotic cells according to the manufacturer’s instructions. Acridine orange was used in the autophagy assay. The cells were stained with 1 μg/mL acridine orange in PBS for 10 min in an incubator. The samples were analyzed by an Attune NxT Flow Cytometer (Thermo Fisher Scientific) and FlowJo software 10.4 (BD Biosciences, Franklin Lakes, NJ, USA).
Apc annexin 5 detection kit with 7 aad
Manufactured by BioLegend
The APC Annexin V detection kit with 7-AAD is a laboratory product used for the detection of apoptosis (programmed cell death) in cell samples. The kit contains Annexin V, a calcium-dependent phospholipid-binding protein, conjugated to the fluorescent dye APC (allophycocyanin). It also includes the nucleic acid dye 7-AAD (7-Aminoactinomycin D).
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3 protocols using apc annexin 5 detection kit with 7 aad
1
Comprehensive Cell Cycle and Apoptosis Analysis
2
Apoptosis and Necrosis Detection by FACS
3
Apoptosis and Necrosis Detection by FACS
Check if the same lab product or an alternative is used in the 5 most similar protocols
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BioLegend APC Annexin V detection kit with 7-AAD (reference 640930) was used to determine apoptosis and necrosis by FACS as follows. 500,000 cells per condition were collected and washed twice in 100ul BioLegend’s cell staining buffer. Cells were then resuspended in Annexin V binding buffer with 5ml of APC Annexin V and 5ml of 7-AAD for 15 minutes in the dark at room temperature. Finally, 400ml of binding buffer was added to each sample and each sample was analyzed by flow cytometry (BD LSRFortessa). At least 100,000 cells were recorded per sample in triplicates. Data were analyzed using FlowJo_V10.
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