Cells were seeded into 6-well plates at 5 × 105 cells/well. After overnight incubation, they were treated with 8 μM DIM for 48 h and medium with same concentrations of DMSO was used as control. After incubation, cells were harvested, then double stained with Annexin V-FITC/PI using an apoptosis analysis kit (KeyGEN BioTECH, CHN), and subjected to flow cytometry analysis for detection of apoptosis. 10,000 cells per sample were analyzed by a BD FACSCalibur Cytometry (BD Biosciences) to quantify apoptotic cells (Annexin V-FITC positive cells).
Apoptosis analysis kit
Manufactured by Keygen Biotech
Sourced in China
The Apoptosis Analysis Kit is a laboratory equipment designed to detect and quantify apoptosis, a programmed cell death process. The kit provides the necessary reagents and protocols to measure various markers associated with apoptosis, enabling researchers to study cell death mechanisms.
Automatically generated - may contain errors
4 protocols using apoptosis analysis kit
1
Apoptosis Analysis of DIM Treated Cells
2
Annexin V-PI Apoptosis Assay
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An apoptosis analysis kit (NanJing KeyGen Biotech Co., Ltd., Nanjing, China) was utilized to detect the cell apoptotic rates. Briefly, the Annexin V and PI were firstly added into the binding buffer, and the treated cells were resuspended in the binding buffer. After incubating in the dark for 15 min, the cells were washed and analyzed using a Guava easyCyte 5 Flow Cytometer (EMD Millipore, Bellerica, MA, USA).
3
Apoptosis Induction in MCF-7 Cells
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MCF-7 cells were seeded into 6-well plates at 5 × 105 cells/well. After overnight incubation, they were pretreated with 20 μM DIM for 2 h and then exposed to 10 Gy of γ-irradiation. Cells were harvested at 48 h after irradiation before being double stained with annexin V-FITC/PI using an apoptosis analysis kit (KeyGEN BioTECH, Nanjing, China) and subjected to flow cytometry analysis for detection of apoptosis. 10,000 cells per sample were analyzed by a BD FACSCalibur Cytometry (BD Biosciences, Franklin Lakes, New Jersey, USA) to quantify apoptotic cells (annexin V-FITC positive cells).
4
Radiosensitivity of MCF-7/ADR Cells
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MCF-7/ADR cells were seeded into six-well plates at 5 Â 10 5 cells/ well. After overnight incubation, they were pretreated with DIM (0, 20, 30 lM) for 2 h and then exposed to 10 Gy of c-irradiation. Cells were harvested at 48 h after c-irradiation before being double stained with Annexin V-FITC/PI using an apoptosis analysis kit (KeyGEN BioTECH) and subjected to flow cytometry analysis for detection of apoptosis. A total of 10,000 cells per sample were analysed by a BD FACSCalibur Cytometry (BD Biosciences) to quantify apoptotic cells (Annexin V-FITC positive cells).
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