Hi All

I'm having some trouble to set up the Proliferation Assay. I'm using Click-IT Edu (Invitrogen) to check proliferation on Jurkat cells.
All my cells are proliferating doesn't matter if it's control, or add with IL-2, or LPS. Same pattern.

Does anyone has some experience with this kit? and these cells?

How long should I put the EdU into the culture?

Thanks a lot

RAFAEL

I too am trying to get this kit to work but with no avail. For $1600 I thought this would be much better!!! I'm having trouble with cells expressing GFP (either control or the tagged construct) not staining. I'm using CHO cells so unfortunately I can't help with the Jurkat cells but I use 10uM EdU for 20 minute pulse before fixation. This was the protocol for BrdU but am now testing 20uM for 20 mins. CHO cells have approximately a 24 hour cell cycle time so in an asynchronous population about 30% are in S phase at any given time. I see this with my non-expressing control but the GFP control (which has been previously shown not to have any deleterious effects on cell cycle) barely stains above non-EdU incorporated control. I'm confused and horribly disappointed. Unless I can get this to work soon I'll have to go back to BrdU.

Thanks for share your experience!!
I was trying put 10uM EdU for 24h. I think it's a long time! I'll try short time for EdU exposition.
Thanks