MC38-OVA and EG7-OVA cells were subjected to 5 or 10 Gy of radiation (or sham-irradiation). After 48 h of incubation in complete medium, the irradiated cells were stained with Annexin V- PE and 7-AAD (eBioscience, USA) according to the manufacturer's instructions. The percentages of apoptotic tumor cells were quantified by flow cytometry. Late apoptotic cells were defined as annexin V and 7-AAD double-positive; early apoptotic cells were defined as annexin V positive but 7-AAD negative.
Annexin 5 pe and 7 aad
Manufactured by Thermo Fisher Scientific
Sourced in United States
Annexin V-PE and 7-AAD are fluorescent dyes used in flow cytometry analysis to assess cell apoptosis. Annexin V-PE binds to phosphatidylserine, which is exposed on the surface of apoptotic cells. 7-AAD is a DNA-binding dye that can penetrate the cell membrane of late apoptotic or necrotic cells. Together, these two dyes can be used to distinguish between viable, early apoptotic, and late apoptotic/necrotic cells.
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Lab products found in correlation
3 protocols using annexin 5 pe and 7 aad
1
Apoptosis Assessment of Irradiated Tumor Cells
2
Annexin V-PE and 7-AAD Apoptosis Assay
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To assess apoptosis, cells were harvested by centrifugation at 1,000 rpm for 5 min, washed twice with cold PBS, resuspended in 500 µl of staining buffer, and co-stained with Annexin V-PE and 7-AAD (eBioscience, San Diego, CA, USA) at room temperature for 15 min in the dark. The cells were analyzed using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA). The data were analyzed using FCS Express software (De Novo Software, Pasadena, CA, USA).
3
Apoptosis and Proliferation Assay
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To assess apoptosis, cells were harvested through centrifugation at 1000 rpm for 5 min, washed twice in cold PBS, resuspended in 500 µl staining buffer, and co-stained with Annexin V-PE and 7AAD (eBioscience, CA, USA) at room temperature for 15 min in the dark, and then analysed using a FACS Calibur ow cytometer (BD Biosciences, NJ, USA) in 1 h. For proliferation assay, cells were pre-treated with the kFlour647 Click-iT Flow detection kit (KeyGEN BioTECH, Jiangsu, China) in accordance with the manufacturer's instructions. The data were analysed by the FCS Express software.
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