Timeline of cytokine production
Lindsey Alexander
Friday, 06 March 2009 19:51 UTC
I need to optimize new rt-PCR primers for research use. I don’t have any positive control, so I am trying to make my own. I have treated splenocytes with anti-CD3 antibody for varying amounts of time from 24-96 hours. My genes of interest are IFN-gamma, IL-10, and IL-17. My results so far have not been completely convincing. Can anyone give me an idea of a timeline of producton of these cytokines from T-cells?
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Replies
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Anonymous
T cells generally require longer time periods for activation unlike b cells due to antigen processing and presentation. i suggest you include day 3, 4 and 5
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I have done time course experiments of T-cell activation from after 6 hrs to 72 hrs. I used to achieve optimal levels of inducible protein expressions after 24 hrs of activation.
But I used other costimulatory molecules (ICOS ligand or CD28) with anti-CD3 to activation the T-cells.
I have also used Mercaptoethanol to provide reducing env to help activation of cells.
But I also think anti-CD-3 itself should also work.. how your control cells are look like after activation?
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