T cell proliferation
Posted 19 September 2003 - 01:26 PM
Posted 24 October 2003 - 11:12 AM
miclino, on Sep 19 2003, 01:26 PM, said:
Normally the peak of proliferation for mitogens (Con A, PHA, etc.) is about 2-3 days and for antigens is 4-6 days. It's means that you should take your cultures to harverster at these times... If you left your cultures with ConA with 6 days of incubation, probably all cells was already died.
Doesn't the matter if you incubate with thymidine for 6 hours, 18 hours or even days. The results will be pretty the same...
Try to see the concentration of the tritiated-thymidine that you are using. I use at 1 uCi per well diluted in culture medium (pulsed for 6 hours).
About your controls being so low, I think could be the long time of culture (6 days + 18 hs). Try to reduce your time of incubation.