Hello.
I am planning an experiments to pulse-label cells with EdU followed by Click reaction and analysis by flow cytometry.
When I performed similar experiments previously, I used to finish the Click reaction in the same day, and kept the cells in the fridge for further analysis.
This time, I need to treat the cells with some drugs for a few hours before EdU labelling, and I am looking for a pausing point.
At the moment, I am thinking of keeping the cells in the fridge after the fixation step (in BSA/PBS).
Alternatively, I may permeabilise the cells and keep in the permeabilisation buffer.
Do you have any comments which is better, or other pausing points that could be better than these two possibilities?
Thanks,
Personally, I would store the cells after fixation. A prolonged incubation in the permeabilization buffer might disrupt the cells too much. However, I have not tried this.
I second Inger. Store the cells in PBS after fixation, but not in the permeabilisation buffer!
Dear Inger and Sushmita,
Thank you very much for your comments/suggestions.
I will write the protocol of my experiment including your points.
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