| | Re: Splenocyte Proliferation Assay
may not be any help, as I have only performed one of these assays (and it was pretty dodgy- I'm only an honours student!!) But, I did get a detectable proliferative response. (my splenocytes were from BALB/c mice).
The media formulation I used was RPMI 500 mL, 2-mercaptoethanol 0.1mM, HEPES 10 mM, Non-essential amino acids 1%, Sodium pyruvate 0.1 mM, L-glutamine 0.02 mM, FCS 10% (and media antibiotics).
I didn't actually use the thymidine method for measuring proliferation though, I used an MTT tetrazolium salt colourimetric assay. This involves adding MTT to each well and incubating for 2 hours, then adding an extraction buffer, allowing colour to develop overnight and reading the OD using an ELISA plate reader.
You probably won't want to change your thymidine method for this one though?? I'm not sure but I think your way is more sensitive?? Anyways, let me know if you want to give it a go and I could give you a protocol.