Problems in T cell proliferation assay

USER USER at elsevier.nl
Fri Dec 29 09:08:34 EST 1995


"Maia Kuprianova" <maia at ibch.siobc.ras.ru> wrote:
>
>Hello,
>I am trying to do a T cell proliferation assay. Although T cells are growing, there 
>is no radioactivity count after addition of 3H-thymidine. Why? Can the specific activity 
>of 3H-thymidine make a difference and what is the optimum of it? Thank you very 
>much.
>
>Maia Kuprianova
>maia at ibch.siobc.ras.ru
>

Hi Maia,

I may be of help to you but I need more info:


1. What kind of cells are these? Mouse or human, peripheral blood or otherwise?

2. What kind of stimulus do you give them? Antigen, PHA?

3. How many cells do you put in what kind of wells (flatbottom, u-bottom?)?

4. Which media and sera do you use?

5. Which 3H-Thymidine do you use and how many microcurie in how much volume?

6. How many days after the initial stimulus do you add the thymidine?

7. How many hours after pulsing with thymidine do you harvest the cells?

8. How do you harvest and count and are you sure the counter is OK?


Please contact me at: d.orsini at elsevier.nl

or post through this newsgroup, but do not send a reply directly, as it will
end up with USER, who is our postmaster.

Daniela




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