I am trying to do aniPOND to selectively capture my protein of interest associated at the replication fork in native condition. For this, I am labeling the replication fork with EdU followed by Click reaction with Biotin azide. I am using double thymidine blocked HeLa S3 (adhering culture) cells. According to the published protocol, I have used 10 uM EdU for 10 minutes pulse. Whereas, I also see some procedures with higher concentrations of EdU (100 uM) or in some cases, 10 uM Edu for 60 minutes pulse. Has anybody performed this experiment in HeLa S3? What EdU concentration and pulse time did you use?
Kulvir Singh
Dear Mandal
The link may be useful to u.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5271410/
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