Help with 3H Thymidine Assay with lymphocytes / cell harvester required?

David Liu via methods%40net.bio.net (by amanosz from yahoo.com)
Tue Oct 2 14:40:45 EST 2007


Hello,

I will be doing a 3H Thymidine uptake assay on primary
T cells for the first time.  The protocol that was
given to me was for an adherent cell line, and I'm
finding that many people use a vacuum cell harvester
to wash non-adherent cells in the 3H thymidine assay. 
Unfortunately, it is difficult for me to have access
to such an instrument, and I'm wondering if it is
possible to do this assay without a vacuum cell
harvester.  Does anyone have any experience with
performing this assay on non-adherent cells without a
harvester?  Is it possible to just centrifuge and
pipette out supernatant to wash away unincorporated
thymidine and TCA?  What recommendations do you have?
Or is a cell harvester a must?

Thanks,
David 



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