In orded to analyze of apoptosis, cells were first cultured in non-serum medium and harvested after 48 h of treatment. Subsequently, the cells were resuspended to 1-5×106/mL by Annexin V binding buffer, and double stained with V-FITC and propidium iodide (BD Biosciences, Franklin Lakes, NJ, USA). Finally, cell apoptosis was analyzed by flow cytometry.
V fitc
Manufactured by BD
Sourced in United States
V-FITC is a fluorescent dye used for labeling and detection in laboratory applications. It emits green fluorescence when excited by a suitable light source, allowing for visualization and analysis of target molecules or cellular structures.
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Lab products found in correlation
4 protocols using v fitc
1
Apoptosis Analysis by Flow Cytometry
2
Apoptosis assessment of intestinal cells
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1 × 106 human intestinal epithelial cells were cultured in a six-well plate and treated with TNF-α and then shRIP3 plasmid or RIP3 inhibitor GSK-872 for 24 h. The cells were incubated with 5 μl V-FITC and 5 μl PI (BD Biosciences Pharmingen, CA, USA) for 15 min in the dark and then resuspend in 400 μl 1 × binding buffer. The proportion of apoptotic cells was analyzed by flow cytometer (Becton, Dickinson and Company, NJ, USA). FlowJo 7.6 software was applied for data analysis.
3
Apoptosis Quantification Protocol
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After 48 h of transfection, cells were trypsinized and collected by the Annexin V binding buffer. Subsequently, cells were double stained with V-FITC and propidium iodide (BD Biosciences, Franklin Lakes, NJ, USA), then measured by BD FACS Canto II (BD Biosciences). Data was analyzed using FlowJo software.
4
Apoptosis Analysis of Transfected PC Cells
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PC cells were transfected with designated sequences for 48 h. Then, cells were harvested, centrifuged, and resuspended with 500 µL of 1X binding buffer (BD Biosciences, Franklin Lakes, NJ, USA). After incubation with 5 µL V-FITC and 5 µL propidium iodide (BD Biosciences, Franklin Lakes, NJ, USA) for 5 min at room temperature in the dark, the apoptosis rate was analyzed by ow cytometry using a Cytomics FC 500 ow cytometer system (Beckman Coulter, Brea, CA, USA) as previously described [12] .
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