Cells were treated with different concentrations of Zoledronate for 3 days, then the supernatant and adherent cells were collected for cell apoptosis analysis using centrifugation at 1,500 × g for 10 min at 4°C. The BD Pharmingen™ APC Annexin V Apoptosis Detection kit (BD Biosciences, Franklin Lakes, NJ, USA) was used according to the manufacturer's instructions. Cells were washed twice with PBS and then suspended in 500 µl binding buffer, 5 µl Annexin V-APC and 1 µl propidium iodide (PI) solution were added into the binding buffer and incubated for 15 min in the dark at 37°C. Cell apoptosis was analyzed by flow cytometry.
Pharmingen apc annexin 5 apoptosis detection kit
Manufactured by BD
Sourced in United States
The BD Pharmingen™ APC Annexin V Apoptosis Detection kit is a laboratory product used for the detection and quantification of apoptosis in cells. It contains Annexin V conjugated with the fluorescent dye allophycocyanin (APC) to enable the identification of apoptotic cells through flow cytometry analysis.
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2 protocols using pharmingen apc annexin 5 apoptosis detection kit
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Zoledronate-Induced Apoptosis Evaluation
2
Annexin V-APC Apoptosis Assay
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Cells were seeded on 6-well plates at a density of 1 × 105 cells/well and then cultured for 72 h. The supernatant and adherent cells were collected and then centrifuged at 1500 rpm for 10 min at 4 °C. For detection of cell apoptosis, the BD Pharmingen™ APC Annexin V Apoptosis Detection kit (BD Biosciences, Franklin Lakes, NJ, USA) was used according to the manufacturer’s protocol. Briefly, the cells were washed with cold PBS twice and resuspended in 500 μL of binding buffer. Then, 5 μL of Annexin V-APC solution was added to the solution and incubated in the dark for 15 min at 37 °C. Finally, 1 μL of energy-coupled dye (ECD-A) solution was added for the subsequent flow cytometry for cell apoptosis analysis.
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