The proliferative capacity of the transfected cell was assessed using the bromodeoxyuridine (Brdu) incorporation assay kits (Millipore, Billerica, MA, USA). Briefly, cells (1×104 (link)) were seeded in each well of a 96-well plate after 48 hours of transfection. 10 µL of Brdu was subsequently added to each well. After 5 hours incubation, cells were fixed by FixDenat for 30 minutes. Cells were then incubated with anti-Brdu antibody in incubation buffer from Roche for 1 hour at 37°C. Cell nuclei were stained with 1 µM SYTOX Green (S7020, Life Technologies) at room temperature for 15 minutes. In each treatment, cells were washed with PBS at least three times.
Protocol detail
Cell Proliferation Assay with Brdu
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Corresponding Organization :
Other organizations : Hubei Cancer Hospital, Air Force Medical University, Zhongnan Hospital of Wuhan University, Wuhan University, Yangtze University
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Variable analysis
independent variables
- Transfection of cells
- Proliferative capacity of the transfected cells
- Cell density (1×10^4 cells per well)
- Incubation time (5 hours)
- Staining with SYTOX Green (1 µM)
- Positive control: Not mentioned
- Negative control: Not mentioned
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