[Immunology] T-cell proliferation suggestion!

Elena Sartorius via immuno%40net.bio.net (by hotsciencegeek from live.com)
Tue Mar 4 18:24:42 EST 2008

Hi N K,With regards to the proliferation assay, i would suggest the following -1. for the cd3 concentration, try 25mg/ml. i find this is highly effective2. instead of using DMEM based media, try using human placental blood. while there maybe ethical issues..data is data :)!3. increase your incubation time to one week.By the way, i have realised that there are not many people in this field - specifically in T-cell proliferation in mice.Would you like to meet for a drink sometime and talk science or more?? with warm regards,Elena Sartorius  Im trying to do a mouse T cell proliferation assay. According to the
literature out there one should be able to induce CD4+ T cell proliferation
by using anti cd3 and anti cd28. But I cant seem to get this to work...i.e.
the cells are not multiplying (im measuring proliferation by using Brdu and
FACS). I have tried different concentrations for cd3 (from 3ug/ml to 25
ug/ml) and cd28 (2ug/ml to 8ug/ml). I also tried them as both platebound and
soluble antibodies. I dont know what I am doing wrong...any help/suggestions
or protocols would be greatly appreciated!

Thank You
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