Click-iT™ Plus EdU Alexa Fluor™ 488 Flow Cytometry Assay Kit

Protocol tips

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	- Cells were stained and incubated for 45 min at 37 °C with 10 μg/ml of propidium iodide and 5 μg/ml of ribonuclease A in PBS.

Protocol tips
- Cells were stained and incubated for 45 min at 37 °C with 10 μg/ml of propidium iodide and 5 μg/ml of ribonuclease A in PBS.

Publication protocol

Flow cytometry was performed using FACS Verse Cell Sorter (BD Biosciences) and analyzed using FlowJo software. For Annexin V staining, cells were labeled with Alexa Fluor 488‐conjugated Annexin V for 15 min in Annexin V‐binding buffer according to the manufacturer's instructions (Invitrogen). For cell cycle analysis, cells were fixed with 70% ice‐cold ethanol and incubated 30 min on ice. Next, cells were incubated for 45 min at 37 °C with 10 μg/ml of propidium iodide and 5 μg/ml of ribonuclease A in PBS. Measurement of ROS and 8‐oxo‐2'‐deoxyguanosine levels was measured as described previously (Venere et al., 2014).

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