Hi all,
I am working on EdU prolifetration assay with flow cytometry. I followed the manufacturer's protocol, however I had a very high signal in negative staining control (with Click-iT but without EdU). In addition, I didn't get two distinctive peaks (figure attached). Does anybody have solutions? Thanks!
Dear Fulei Wuchu
I have never used that kit, but I have done cell cycle analysis using Hoechst 33342.
In that case, I had to use a linear scale to display the histogram, not a log scale. You may want to change your diagram to a linear scale as well.
Best,
I think the idea of Dr. Narumi Uno may give good results
All the best,
Hany
Dear All,
It is important and highly recommended to use Click-iT Alexa 647 but not Alexa 488 due to autoflourescence induced high background. Actually, I do not know which one you are using.
Good luck
Imam
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